Δ2,3 -0-2-Isocephem-4-carboxylic acid and derivatives as antibacterial agents

ABSTRACT

There is described the stereoselective total synthesis of novel Δ 2 ,3 -1,4-morpholine-2-carboxylic acids possessing a fused β-lactam ring in the 1,6-position and carrying a substituent cis to carbon 5 in the 7-position of the fused ring system represented by the general formula ##STR1## wherein X is amino and Z represents an optionally substituted heterocyclic ring containing N, O or S. Also included in the invention are compounds of formula I in which the carboxyl group at the 2-position is protected as by an easily cleavable ester group and salts of both the free acids and carboxyl-protected compounds of formula I. The compounds of formula I are potent antibacterial agents or are of use as intermediates in the preparation of such anti-bacterial agents.

BACKGROUND OF THE INVENTION

1. Field of the Invention

The chemical processes of the present invention produce novelantibacterial agents of the β-lactam type containing a hitherto unknownnucleus and useful intermediates for their synthesis.

2. Description of the Prior Art

Penicillins and cephalosporins comprise a group of well-knownantibacterial agents commonly grouped together as a class calledβ-lactam antibiotics. For a recent review of this field with manycitations (especially the first ten) to the prior work, see J. P. Houand J. W. Poole, β-lactam Antibiotics: Their Physiocochemical Propertiesand Biological Activities in Relation to Structure, J. PharmaceuticalSciences, 60(4), 503-532 (April, 1971). Most of the work in this fieldhas fundamentally been done, speaking broadly, with 6-aminopenicillanicacid, 7-aminocephalosporanic acid and derivatives thereof produced byfermentation or chemical transformation of the natural products. Despitethe extensive progress made in preparing active derivatives of6-aminopenicillanic acid and 7-aminocephalosporanic acid, there is acontinuing search for synthetic and semi-synthetic routes to newfamilies of β-lactam antibiotics which may possess more advantageousproperties then those derived from the known pencillin and cephalosporinnuclei.

Considerable work has been done on total chemical synthesis of bothknown β-lactams and nuclear analogs of such known compounds. A recentreview is the text by M. S. Manhas and A. K. Bose, Synthesis ofPenicillin, Cephalosporin C and Analogues, Marcel Decker, Inc., 95Madison Avenue, New York, New York, 1969. Another extensive review is byR. B. Morin and B. G. Jackson, Chemistry of Cephalosporin Antibiotics,Fortschr. Chem. Org. Naturst., 28, 343-403 (1970), especially pages379-393; the now famous "Woodward Intermediate" is shown therein ascompound 146 on page 387. A more recent review of β-lactams is that byM. S. Manhas and A. K. Bose, Beta-Lactams: Natural and Synthetic: Part1, Wiley-Interscience, New York, New York, 1971. A still further reviewarticle on the synthesis of β-lactams is that by A. K. Mukerjee et al.,Synthesis, 327 (1973).

Other pertinent publications relating to synthesis of β-lactams are:

a. D. M. Brunwin, G. Lowe and J. Parker, J.C.S. Chem. Comm., 1971,865-867, describing synthesis of nuclear analogs of thepenicillin-cephalosporin group.

b. D. M. Brunwin et al., J. Chem. Soc. (C), 1971, 3756-3762 and J.C.S.Chem. Comm., 1972, 589-590 describing total synthesis of nuclear analogsof penicillins and cephalosporins.

c. S. Kukolja, J. Amer. Chem. Soc., 93, 6267-6270 (1971) and 94,7590-7593 (1972) describing preparation of6-phthalimido-5-epipenicillanates and disulfide analogs of penicillins.

d. J. A. Webber et al., J. Medicinal Chemistry, 14(11), 1136-1138 (1971)describing preparation of 3-cyanomethyl cephem nucleus.

e. West German Patent Specification 2,219,601 (Farmdoc 76,051T)describing synthesis of β-lactams of the formula ##STR2## wherein X ishalo, N₃ -- or H₂ N--, A is --S--, --S--CH₂ --, --o--, --O--CH₂ --,--CH₂, --CH₂ CH₂ -- or --NH-- and R¹, R² and R³ are hydrogen, C₁ --C₆alkyl or aryl.

f. U.K. Patent 1,308,822 disclosing β-lactams of the formula ##STR3##where Y = amino or substituted amino.

g. S. Wolfe et al., Can. J. Chem., 50, 2894-2905 (1972) describingsynthesis of sulfur-free penicillin derivatives.

h. French Patent 2,111,859 describing nuclei of the formula ##STR4## and7-acylated derivatives thereof.

i. Helvetica Chimica Acta, 55(2), 388-429 (1972) describing nuclearmodified cephalosporins and penicillins.

j. F. Moll et al., Zeit. fur Naturforsch. B, 27(b)6, 727 (1972)describing nuclear analogs of cephalosporins.

k. U.K. Specifications Nos. 1,271,013 and 1,271,014 describing γ-lactamsof 7-(acylamino)-3-aminomethyl-ceph-3-em-4-carboxylic acids.

l. U.K. Specification 1,271,180 describing preparation of novelthiazoline azetidinone rearrangement products useful as intermediates inpenicillin and cephalosporin synthesis.

m. German Patent Specifications No. 2,046,822, 2,046,823 and 2,046,824describing synthesis of novel azetidinone intermediates.

n. G. Lowe et al., J. Chem. Soc. Perkins I, 1322 (1973) describing totalsynthesis of nuclear analogs of 7-methylcephalosporins having theformula ##STR5##

o. D. M. Brunwin et al., J. Chem. Soc. Chem. Comm., 865 (1971)describing synthesis of compounds of the formula ##STR6##

S. Wolfe et al., Canadian J. Chem., 50, 2902 (1972) describing compoundsof the formula ##STR7##

q. J. P. Luttringer et al., Tetrahedron Letters, 4163-4166 (1973)describing compounds of the formula ##STR8##

r. U.S. Pat. No. 3,835,130 disclosing β-lactams of the formula ##STR9##wherein R_(a) and R_(b) are each hydrogen or R_(a) and R_(b) form acovalent carbon-to-carbon bond, R₁ ' represents hydrogen, R₁ " iscyanoacetyl, bromacetyl or an acyl group of the formula ##STR10## inwhich Ar is phenyl, 3-hydroxyphenyl, 4-hydroxyphenyl,3-chloro-4-hydroxyphenyl, 3,5-dichloro-4-hydroxyphenyl or 2-thienyl, R₂' is hydrogen and R₃ ' is hydrogen or lower alkyl.

s. West German Offenlegungsschrift 2,355,209 describing synthesis ofβ-lactams of the formula ##STR11## wherein R is acyl, A' is hydrogen,hydroxy, carbamoyloxy, thiocarbamoyloxy, quaternary ammonium, N-loweralkyl carbamoyloxy, N,N-di-lower alkyl carbamoyloxy, N-lower alkylthioand N,N-diloweralkylthiocarbamoyloxy, azido, halo, cyano, a tertiaryamine, acyloxy, or a 5-member heterocyclic thio group having 1-4 heteroatoms; B is H, OCH₃ or SR where R is lower alkyl or aryl; X is adivalent radical selected from --O--, --2-- or --NY-- where Y ishydrogen, lower alkyl, formyl or benzyl and R' is hydrogen or aprotecting group.

Replacement of the 3-acetoxy group of a cephalosporin by variousheterocyclic thiols has been extensively discussed in the literature.Issued patents on 3-thiolated cephalosporins in which the 7-substituentis

1. α-Amino-α-phenylacetamido include U.S. Pat. Nos. 3,641,021,3,734,907, 3,687,948, 3,741,965, 3,757,015, 3,743,644, Japan 71/24400(Farmdoc 46374S), Belgium 776,222 (Farmdoc 38983T; U.K. 1,328,340 whichincludes various substituents on the benzene ring), Belgium 772,592(Farmdoc 19696T; U.S. Pat. Nos. 3,687,948, 3,734,907 and 3,757,012),West Germany 2,202,274 (Farmdoc 50428T) corresponding to U.S. Pat. No.3,759,904, Netherlands 7205644 (Farmdoc 76309T; U.S. Pat. Nos.3,757,014); 3,855,213 and 3,867,380; and

2. o-, m- or p-aminoethoxyphenylacetamido as Netherlands 72/13968(Farmdoc 24740U) corresponding to U.S. Pat. No. 3,759,905 and

3. o-aminomethylphenylacetamido as Netherlands 72/06326 (Farmdoc 76374T)(which also reviews the older patent literature concerning substituted7-phenylacetamidocephalosporanic acids) corresponding to U.S. Pat. No.3,766,176 and 3,766,175; and

4. N-(phenylacetimidoyl)aminoacetamido as U.S. 3,692,779; and

5. α-amino-α-(1,4-cyclohexadienyl)acetamido as in Belgium 776,222(Farmdoc 38983T; U.K. 1,328,340).

Additional similar disclosures are found in U.S. Pat. No. 3,692,779(Belgium 771,189; Farmdoc 12819T), Japan 72/05550 (Farmdoc 12921T),Japan 72/05551 (Farmdoc 12922T), U.S. 3,719,673 (Belgium 759,570;Farmdoc 39819S), Belgium 793,311 (Farmdoc 39702U) and Belgium 793,191(Farmdoc 39684U).

Issued disclosures of 3-thiolated cephalosporins in which the7-substituent is 7-mandelamido (7-α-hydroxyphenylacetamido) are found,for example, in U.S. Pat. No. 3,641,021, France 73.10112, U.S. Pat. No.3,796,801, Great Britain 1,328,340 (Farmdoc 38983T), U.S. 3,701,775,Japan 4844293 (Farmdoc 55334U), U.S. 3,855,213, and in Hoover et al., J.Med. Chem. 17(1), 34-41 (1974) and Wick et al., Antimicrobial Ag.Chemo., 1(3), 221-234 (1972).

U.S. Pat. No. 3,819,623 (and, for example, also U.K. 1,295,841 and WestGermany 1,953,861) discloses specifically and with working details thepreparation of 2-mercapto-1,3,4-thiadiazole-5-acetic acid and itsconversion to7-(1H-tetrazol-1-ylacetamido)-3-(5-carboxymethyl-1,3,4-thiadiazol-2-ylthiomethyl)-3-cephem-4-carboxylicacid which is also disclosed in West Germany Offenlegungsschrift2,262,262.

SUMMARY OF THE INVENTION

The present invention provides steroselective total synthesis of certainnovel substituted Δ²,3 -1, 4-morpholine-2-carboxylic acids possessing afused β-lactam ring in the 1,6-position and carrying a substituent cisto carbon 5 in the 7-position of the fused ring system represented bythe general formula ##STR12## wherein X is amino, azido or acylamido andZ represents optionally substituted C₁ -C₆ alkyl, aryl, aralkyl orheterocyclic. When X is acylamino, these acids (and theirpharmaceutically acceptable salts and physiologically hydrolyzed esters)are potent antibacterial agents.

Also included in this invention are various novel intermediates usefulin preparing the active β-lactam derivatives described above and variousprocesses for the production of the intermediates and active compounds.

The compounds having the above general formula represent a new family ofβ-lactam antibiotics. They can be considered nuclear analogs ofcephalosporins in which the sulphur atom of the dihydrothiazine ring isreplaced by an oxygen atom and shifted from position 5 to position 4 ofthe β-lactam ring system as numbered in the formula above. Thenomenclature to be used could be the following: ##STR13##

However, Sheehan has used the term O-cephem for the structure ##STR14##[J. C. Sheehan and M. Dadic, J. Heterocyclic Chem., 5, 770 (1968)] andwe propose the use of the term 0-2-isocepham for the basic system havingthe formula ##STR15## The numerical prefix indicates the position of thehetero-atom.

To illustrate the above system, the intermediate of the formula##STR16## may be named benzyl7β-amino-3-(1-methyltetrazol-5-ylthiomethyl) -Δ³-0-2-isocephem-4-carboxylate and the compound of the formula ##STR17##may be named7β-(2-aminomethylphenylacetamido)-3-(1-methyltetrazol-5-ylthiomethyl)-Δ³-0-2-isocephem-4-carboxylic acid.

There is thus provided by the present invention the novel 0-2-isocephemcarboxylic acid compounds having the formula ##STR18## wherein R is anacyl group and Z represents optionally substituted C₁ -C₆ alkyl, aryl,aralkyl or heterocyclic, and easily cleavable esters andpharmaceutically acceptable salts of said acids and esters.

The acyl group R can be chosen from a wide variety of organic acylradicals which yield products of improved properties and is preferablyan acyl radical which is contained in a naturally occurring orbiosynthetically, semi-synthetically or totally-synthetically producedpharmacologically active N-acyl derivative of 6-aminopenicillanic acidor 7-aminocephalosporanic acid. Examples of suitable acyl groups aredefined in the following general formulae, but it should be noted thatthis is not intended to be an exhaustive list of all the possible acylgroups which may be used.

i. R^(a) C_(n) H_(2n) CO--

where R^(a) is aryl (carbocyclic or heterocyclic), substituted aryl,cycloalkyl, substituted cycloalkyl, cycloalkenyl, substitutedcycloalkenyl or a nonaromatic or mesoionic heterocyclic group, and n isan integer from 1-4. The preferred R^(a) substituents are (a) arylselected from phenyl, 2-thienyl, 3-thienyl, furyl, 4-isoxazolyl,pyridyl, tetrazolyl, sydnone-3 or -4, imidazolyl, naphthoyl,quinoxalinyl, triazolyl, isothiazolyl, thiadiazolyl, thiazolyl,oxazolyl, oxadiazolyl, pyrazolyl, furazan, pyrazinyl, pyrimidinyl,pyridazinyl or triazinyl; (b) substituted aryl in which the aryl groupsmentioned above under (a) are substituted by one or more radicalsselected from chloro, bromo, iodo, fluoro, nitro, amino, cyano,(lower)alkanoyloxy, (lower)alkanoyl, (lower)alkoxyamino, (lower)alkoxy,(lower)alkyl, (lower)alkylamino, hydroxy, guanidino, (lower)alkylthio,carboxy, phenyl, halophenyl, trifluoromethyl, di(lower)alkylamino,sulfamyl, (lower)alkanoylamino, phenyl(lower)alkylamido,cycloalkylamino, allylamido morpholinocarbonyl, pyrrolidinocarbonyl,piperidinocarbonyl, tetrahydropyridino, furfurylamido orN-alkyl-N-anilino; (c) C₃ -C₁₂ cycloalkyl; (d) substituted C₃ -C₁₂cycloalkyl where the substituents are one or more radicals selected fromchloro, bromo, fluoro, iodo, nitro, trifluoromethyl, C₁ -C₄ alkyl, C₁-C₄ alkylamino, C₁ -C₂ alkoxy or amino; (e) C₃ -C₁₂ cycloalkenyl, saidcycloalkenyl group having 1 or 2 double bonds; and (f) substituted C₃-C₁₂ cycloalkenyl, said cycloalkenyl group having 1 or 2 double bondsand being substituted by one or more radicals selected from chloro,bromo, fluoro, iodo, nitro, trifluoromethyl, C₁ -C₄ alkyl, C₁ -C₄alkylamino, C₁ -C₂ alkoxy or amino. The most preferred R^(a) groups arephenyl; phenyl substituted by one or more radicals selected from chloro,bromo, iodo, fluoro, nitro, amino, (lower)alkyl, guanidino,(lower)alkylthio, cyano, (lower)alkoxy, sulfamyl, (lower)alkylamino,hydroxy, acetoxy, or trifluoromethyl; 2-thienyl; 3-thienyl; tetrazolyl;sydnone -3; sydnone -4; furyl; isothiazolyl; thiadiazolyl optionallysubstituted with phenyl; oxadiazolyl optionally substituted with phenyl;thiazolyl; imidazolyl; triazolyl; oxazolyl; pyridyl; furazan optionallysubstituted at the 3-position with methoxy; 4-isoxazolyl optionallysubstituted at the 5-position with methyl and at the 3-position withphenyl or halophenyl; 1,4-cyclohexadienyl; 1-cyclohexenyl and1-aminocyclohexyl.

The most preferred acyl groups of this category are those in which nis 1. Examples of this category include phenylacetyl, halophenylacetyl,nitrophenylacetyl, aminophenylacetyl, β-(o-aminomethylphenyl)propionyl,(lower)alkanoyloxyphenylacetyl (e.g. p-acetoxyphenylacetyl),(lower)alkoxyphenylacetyl (e.g. methoxyphenylacetyl,ethoxyphenylacetyl), (lower)alkylphenylacetyl (e.g. methylphenylacetylor ethylphenylacetyl), hydroxyphenylacetyl (e.g. o-hydroxyphenylacetyl),(lower)alkylaminophenylacetyl (e.g. o-, m- or p-aminomethylphenylacetyl), o- m- or p- guanidinophenylacetyl,o-carboxyphenylacetyl, N,N-bis-(2-chloroethyl)aminophenylpropionyl,thien-2 and 3-ylacetyl, 2- or 3- furylacetyl,1,2,5-thiadiazole-3-acetyl, isothiazolyl-4-acetyl, 4-isoxazolylacetyl,1-cyclohexenylacetyl, 2-aminomethyl-1-cyclohexenylacetyl,1-aminocyclohexylacetyl, 1,4-cyclohexadienylacetyl,2-aminomethyl-1,4-cyclohexadienylacetyl, pyridylacetyl, tetrazolylacetyl(other heterocyclic groups of this type are disclosed in U.S. Pat. No.3,819,623 and 3,516,997) or a sydnoneacetyl groups as disclosed in U.S.Pat. Nos. 3,681,328, 3,530,123 and 3,563,983. Other groups of this typeinclude 3-phenyl-5-chlorophenyl-5-methylisoxazol-4-ylacetyl and3-(2,6-dichlorophenyl)-5-methylisoxazol-4-ylacetyl or a group in whichisoxazolyl is replaced by isothiazole as disclosed in U.S. 3,551,440.Still other examples are o-, m- and p-(2'-aminoethoxy)phenylacetyl (asdisclosed in U.S. 3,759,905),4,5-dimethoxycarbonyl-1,2,3-triazol-1-ylacetyl or4-cyano-1,2,3-triazol-1-yl-acetyl (as disclosed in U.S. Pat. No.3,821,206) and imidazol-(1)-acetyl (as disclosed in U.S. Pat. No.3,632,810;

ii C_(n) H_(2n) ₊₁ CO--

where n is an integer from 1-7. The alkyl group may be straight orbranched and, if desired, may be interrupted by an oxygen or sulphuratom or substituted by, e.g., a cyano group. Examples of this groupinclude cyanoacetyl, valeryl, hexanoyl, heptanoyl, ethoxycarbonyl,octanoyl and butylthioacetyl. A preferred acyl group is cyanoacetyl;

iii. C_(n) H_(2n) ₋₁ CO--

where n is an integer from 2-7. The alkenyl group may be straight orbranched and, if desired, may be interrupted by an oxygen or sulphuratom. An example of this group is allylthioacetyl; ##STR19## where R^(a)is as defined under (i) and in addition may be benzyl, C₁ -C₆ alkyl or(lower)alkoxycarbonyl and R^(b) and R^(c) which may be the same ordifferent each represent hydrogen, phenyl, benzyl, phenethyl or C₁ -C₆alkyl. The preferred R^(a) substituents in this category are benzyl, C₁-C₆ alkyl, (lower)alkoxycarbonyl and those mentioned under (i) as beingpreferred aryl, substituted aryl, cycloalkyl (and substitutedcycloalkyl) and cycloalkenyl (and substituted cycloalkenyl) groups. Themost preferred R^(a) group is phenyl. Examples of this group includephenoxyacetyl, 2-phenoxy-2-phenylacetyl, 2-phenoxypropionyl,2-phenoxybutyryl, benzyloxyacetyl, 2-methyl-2-phenoxypropionyl,p-cresoxyacetyl, p-methylthiophenoxyacetyl and ethoxycarbonylacetyl;##STR20## where R^(a) is as defined under (i) and in addition may bebenzyl or C₁ -C₆ alkyl and R^(b) and R^(c) have the meanings definedunder (iv). The preferred R^(a) substituents in this category arebenzyl, C₁ -C₆ alkyl and those mentioned under (i) as being preferredaryl, substituted aryl, cycloalkyl (and substituted cycloalkyl) andcycloalkenyl (and substituted cycloalkenyl) groups. The most preferredaryl groups of this type are those in which R^(b) and R^(c) are hydrogenand R^(a) is phenyl; phenyl substituted with one or more radicalsselected from chloro, bromo, iodo, fluoro, nitro, amino, (lower)alkyl,(lower)alkylthio, cyano, (lower)alkoxy, (lower) alkylamino, hydroxy,acetoxy or trifluoromethyl; 3-pyridyl; or 4-pyridyl;

vi. R^(a) X (CH₂)_(m) CO--

where R^(a) is as defined under (i) and in addition may be benzyl, X isoxygen or sulphur and m is an integer from 2-5. The preferred R^(a)groups are benzyl and those mentioned under (i) as being preferred aryl,substituted aryl, cycloalkyl (and substituted cycloalkyl) andcycloalkenyl (and substituted cycloalkenyl) groups. An example of thisgroup is S-benzylthiopropionyl.

vii. R^(a) CO--

where R^(a) is as defined under (i). The preferred R^(a) groups arethose mentioned under (i) as being preferred aryl, substituted aryl,cycloalkyl (and substituted cycloalkyl) and cycloalkenyl (andsubstituted cycloalkenyl) groups. The most preferred aryl groups of thiscategory are those in which R^(a) is phenyl; phenyl substituted with oneor more radicals selected from chloro, bromo, iodo, fluoro, nitro,amino, (lower)alkyl, (lower)alkylthio, cyano, (lower)alkoxy,(lower)alkylamino, di(lower)alkylamino, hydroxy, acetoxy ortrifluoromethyl, and most preferably phenyl substituted at the2-position by carboxy or phenyl or at the 2- and 6-positions by methoxy;2-ethoxynaphthoyl; 3-phenyl-5-methylisoxazol-4-yl;3-o-chlorophenyl-5-methylisoxazol-4-yl;3-(2,6-dichlorophenyl)-5-methylisoxazol-4-yl and 1-aminocyclohexyl.Examples of this group include 2,6dimethoxybenzoyl, benzoyl,2-biphenylcarbonyl, 2-aminomethylbenzoyl,2-carboxybenzoyl-2-phenylbenzoyl, 2-thienylcarbonyl, 3-thienylcarbonyland 2-chlorobenzoyl; ##STR21## where R^(a) is as defined under (i) and Yis hydrazino, guanidino, ureido, thioureido and substituted thioureido(as disclosed in U.S. Pat. No. 3,741,962), allophanamido (as describedin U.S. Pat. No. 3,483,188), 3-guanyl-1-ureido (as in U.S. Pat. No.3,796,709), 3-(2-furoyl)ureido, cyanamino (as in U.S. Pat. No.3,796,709), 3-(benzoyl)ureido, azido, amino, acylamino (e.g.carbobenzoxyamino), a group obtained by reacting the amino group of the7-side chain with an aldehyde or ketone (e.g. acetone, formaldehyde,acetaldehyde, butyraldehyde, acetylacetone, methyl acetoacetate,benzaldehyde, salicylaldehyde, methyl ethyl ketone or ethylacetoacetate), hydroxy, etherified hydroxy, esterified hydroxy, carboxy,esterified carboxy (as disclosed for example in U.S. Pat. Nos.3,282,926, 3,819,601 and 3,635,961 and including especially ##STR22##tetrazolyl, cyano, halogeno, acyloxy (e.g. formyloxy or(lower)alkanoyloxy), sulfo, sulfoamino or esterified sulfo. Thepreferred R^(a) substituents are those mentioned under (i) as beingpreferred aryl, substituted aryl, cycloalkyl (and substitutedcycloalkyl) and cycloalkenyl (and substituted cycloalkenyl) groups.Preferred Y substituents are hydrazino; guanidino; ureido; substitutedthioureido of the formula ##STR23## in which R^(p) is hydrogen or C₁ -C₈alkyl and R^(q) is hydrogen, C₁ -C₈ alkyl, C₂ -C₈ alkenyl, phenyl,benzoyl, C₁ -C₈ alkoxy- C₁ -C₈ alkyl, (carbo-C₁ -C₈ alkoxy) C₁ -C₈alkyl; allophanamido; 3-guanyl-1-ureido; 3-(2-furoyl)ureido;3-(benzoyl)ureido; azido; amino; a group obtained by reacting the aminogroup Y with acetone, formaldehyde, acetaldehyde, butyraldehyde,acetylacetone, methyl acetoacetate, benzaldehyde, salicylaldehyde,methyl ethyl ketone or ethyl acetoacetate; hydroxy; etherified hydroxyincluding especially (lower)alkoxy; carboxy; esterified carboxyincluding especially 5-indanyloxycarbonyl; triazolyl; tetrazolyl; cyano;cyanamino; halogeno; formyloxy; (lower)alkanoyloxy; sulfo; orsulfoamino. Examples of this group include α-aminophenylacetyl;α-carboxyphenylacetyl; 2,2-dimethyl-5-oxo4-phenyl-1-imidazolyl;α-amino-p-hydroxyphenylacetyl; α-hydroxyphenylacetyl;α-formyloxyphenylacetyl and other aryl groups of this type disclosed inU.S. Pat. Nos. 3,812,116 and 3,821,017; α-amino-α-2- or 3-thienylacetyl;α-amino-α-(3-chloro-4-hydroxy)phenylacetyl;α-amino-α-(1,4-cyclohexadienyl)acetyl; α-azidophenylacetyl;α-amino-α-(1-cyclohexenyl)acetyl; 2-carboxy-α-3-thienylacetyl;α-amino-α-(3,5-dichloro-4-hydroxyphenyl)acetyl; α-amino-α-3- or 4- or5-isothiazolylacetyl (as in U.S. Pat. No. 3,579,506) and other α-aminoand α-hydroxy-heterocyclylacetyl groups as disclosed for example in U.S.Pat. No. 3,821,207; ##STR24## where R^(d), R^(e) and R^(f) which may bethe same or different may each represent C₁ -C₆ alkyl, phenyl orsubstituted phenyl. The preferred phenyl substituents are one or moreradicals selected from chloro, bromo, iodo, fluoro, trifluoromethyl,nitro, amino, cyano, (lower)alkanoyloxy, (lower)alkanoyl,(lower)alkoxyamino, (lower)alkoxy, (lower)alkyl, (lower)alkylamino,hydroxy, (lower)alkylthio, carboxy di(lower)alkylamino or sulfamyl. Anexample of this group is triphenylmethylcarbonyl. ##STR25## where R^(a)is as defined under (i) and in addition may be hydrogen, C₁ -C₆ alkyl,halogen substituted C₁ -C₆ alkyl, phenethyl, phenoxymethyl; benzyl or##STR26## and X is oxygen or sulphur. An example of such a group isCl(CH₂)₂ NHCO; ##STR27## where Y is as defined under (viii) and n is aninteger of 1-4. A most preferred Y substituent is amino. An example ofthis group is 1-aminocyclohexanecarbonyl.

xii. Aminoacyl, for example

R^(g) CH(NH₂) -(CH₂)_(n) CO-

where n is an integer of 1-10, or

H₂ N-C_(n) H_(2n) Ar(CH₂)_(m) Co-

where m is zero or an integer from 1-10, and n is 0, 1, or 2; R^(g) ishydrogen or an alkyl, aryl, aralkyl or carboxy group or a group asdefined under R^(a) in (i) above; and Ar is an arylene group, e.g.p-phenylene or 1,4-naphthylene. Preferred aryl groups of the aboveformulae are those in which R^(g) is hydrogen, (lower)alkyl, phenyl,benzyl or carboxy and Ar is p-phenylene or 1,4-naphthylene. Examples ofsuch groups are disclosed in U.K. Patent No. 1,054,806. Examples ofgroups of this type include p-aminophenylacetyl and δ-aminoadipoylderived from naturally occurring amino acids and derivatives thereof,e.g. N-benzoyl-δ-aminodipoyl;

xiii. Substituted glyoxylyl groups of the formula

R^(h). CO.sup.. CO-

where R^(h) is an aliphatic, araliphatic or aromatic group. Thepreferred R^(h) groups are 2-thienyl; 3-thienyl; α-naphthyl;2-phenanthryl or a mono-, di- or trisubstituted phenyl group, thesubstituents being selected from chloro, bromo, iodo, fluoro, amino,di(lower)alkylamino, (lower)alkyl, (lower)alkoxy, nitro or(lower)alkanoylamino. Examples of this category are disclosed in U.S.Pat. Nos. 3,546,219 and 3,573,294. Included in this group are also theα-carbonyl derivatives of the above substituted glyoxylyl groups formedfor example with hydroxylamine, semicarbazide, thiosemicarbazide,isoniazide or hydrazine; ##STR28## where R^(a) has the meaning definedunder (i), X is oxygen or sulphur, X' is oxygen or imino and R^(i)represents (lower)alkyl, cycloalkyl having 4,5, 6 or 7 carbon atoms,monohalo(lower)alkyl, dichloromethyl, trichloromethyl, (lower)alkenyl of2-6 carbon atoms, ##STR29## n is an integer from 0 to 3 inclusive andeach of R^(k) and R^(j) is hydrogen, nitro, di(lower)alkylamino,(lower)alkanoylamino, (lower)alkanoyloxy, C₁ -C₆ alkyl, C₁ -C₆ alkoxy,sulfamyl, chloro bromo, iodo, fluoro or trifluoromethyl. The preferredR^(a) substituents are those mentioned under (i) as being preferredaryl, substituted aryl, cycloalkyl (and substituted cycloalkyl) andcycloalkenyl (and substituted cycloalkenyl)groups. Preferred acyl groupsof this type are those in which R^(a) is 2-thienyl; 3-thienyl; phenyl;or phenyl substituted by one or more radicals selected from nitro,di(lower)alkylamino, (lower)alkanoylamino, amino, hydroxy,(lower)alkanoyloxy, C₁ -C₆ alkyl, C₁ -C₆ alkoxy, sulfamyl, chloro,bromo, iodo, fluoro or trifluoromethyl; X is oxygen; X' is oxygen orimino and R^(i) is (lower)alkyl, phenyl, 2-thienyl, 3-thienyl, 2-furylor 5-nitro-2-furyl. The most preferred groups are those of the aboveformula where R^(a) is phenyl, p-hydroxyphenyl, 2-thienyl or 3-thienyl;X is oxygen; X' is oxygen, and R.sup. i is phenyl or 2-furyl. Examplesare disclosed in U.S. Pat. Nos. 3,687,949 and 3,646,024; ##STR30## whereR^(a) has the meaning defined in (i) and R^(i) has the meaning definedin (xiv). The preferred R^(a) substituents are those mentioned under (i)as being preferred aryl, substituted aryl, cycloalkyl (and substitutedcycloalkyl) and cycloalkenyl (and substituted cycloalkenyl) groups.Preferred R^(i) substituents include (lower)alkyl, dichloromethyl, C₄-C₇ cycloalkyl, 2-thienyl, 3-thienyl, phenyl, benzyl, halobenzyl,##STR31## Examples of this group are disclosed in U.S. Pat. Nos.3,626,024 and 3,692,779; ##STR32## where R^(a) has the meaning definedin (i) and R¹ is (lower)alkyl, C₃ -C₁₂ cycloalkyl, aryl (especiallyphenyl), a monocyclic heterocyclic radical having 5 or 6 atoms exclusiveof hydrogen which are C, S, N or O, no more than 2 atoms being otherthan C, or a substituted monocyclic heterocyclic radical as definedabove having one or more substituents selected from halo, (lower)alkyl,(lower)alkoxy or phenyl. Examples of this group are disclosed in U.S.Pat. No. 3,778,436. Most preferred R¹ groups are (lower)alkyl, phenyl,thienyl or furyl.

A preferred class of acyl groups are those of the formula ##STR33##wherein Ar' is a radical of the formula ##STR34## in which R^(m), R^(n)and R^(o) are alike or different and each is hydrogen, hydroxy,(lower)alkyl, cyano, (lower)alkoxy, chloro, bromo, iodo, fluoro,trifluoromethyl, nitro, amino, (lower)alkylamino, di(lower)alkylamino,(lower)alkanoyl, (lower)alkanoyloxy such as p-acetoxy or phenyl and Y isamino or a group obtained by reacting the amino group with acetaldehyde,formaldehyde or acetone, fluoro, chloro, bromo, iodo, hydroxy,(lower)alkanoyloxy, carboxy, guanidino, 3-guanyl-1-ureido,3-(2-furoyl)ureido, 3-benzoylureido, sulfo, sulfoamino, ureido,thioureido, (lower)alkoxy, cyano, cyanamino or indanyloxycarbonyl.Particularly preferred Ar radicals are phenyl, p-hydroxyphenyl,4-hydroxy-3,5-dichlorophenyl, 3-chloro-4-hydroxyphenyl, o-, m- or p-aminomethylphenyl, 2-thienyl, 3-thienyl, 1-cyclohexenyl and1,4-cyclohexadienyl. Particularly preferred Y groups are amino, hydroxyand carboxy. Set forth below are formulae of the most preferred acylgroups of this class: ##STR35## Of most interest are the acyl groups ofthe above class where the acid ArCH(X)COOH is of the D-series.

Other particularly preferred acyl groups for the compounds of formula Iare ##STR36## where U and V are alike or different and each is hydrogen,chloro or fluoro; ##STR37##

Substituent Z in formulae I and II above may be C₁ -C₆ alkyl, aryl,aralkyl or heterocyclic, any of said groups being optionally substitutedby one or more substituents. The alkyl group may be a monovalentsaturated aliphatic hydrocarbon radical having from 1 to 6 carbon atomsand a straight or branched chain, e.g., methyl, ethyl, propyl,isopropyl, butyl, isobutyl, t-butyl, amyl or hexyl. The aryl group maybe a mono-, bi- or polycyclic aromatic hydrocarbon radical, e.g.,phenyl, 1-naphthyl, 2-naphthyl or 2 phenanthryl. The term aralkyl asused herein includes monovalent aryl-substituted aliphatic hydrocarbonradicals of the formula aryl-(ALK)_(m) -- in which aryl is as definedabove, m is an integer of 1 to 4 and ALK represents a straight orbranched chain alkylene radical, e.g., -(ALK)_(m) -- may be methylene,ethylene, propylene, butylene, 1-methylpropylene, 2-ethylethylene andthe like. Heterocyclic Z substituents may be heteromonocyclic orheterobicyclic residues of aromatic character as well as appropriatepartially or wholly saturated residues.

A preferred group of compounds of formulae I and II are those in which Zis C₁ -C₆ alkyl, aryl selected from phenyl or naphthyl, aralkyl of theformula aryl-(ALK)_(m) -- in which aryl is phenyl or naphthyl, m is aninteger of 1 to 4 and ALK represents a straight or branched chainalkylene radical, or heterocyclic selected from a 5- or 6- memberedheterocyclic ring containing 1 to 4 atoms selected from N, O or S, saidalkyl radical being optionally substituted by one or more substituentsselected from hydroxy, halo, amino, nitro, di(C₁ -C₄ alkyl) amino,carboxy, sulfo or cyano and said aryl, aralkyl or heterocyclic radicalsbeing optionally substituted by one or more substituents selected fromhalo, C₁ -C₄ alkyl, C₁ -C₄ alkoxy, cyano, carboxyl, amino, nitro, C₃ -C₄cycloalkyl, C₂ -C₄ alkenyl, trifluoromethyl, hydroxy, hydroxymethyl, C₁-C₄ alkylthio, C₁ -C₄ alkylamino, di(C₁ -C₄ alkyl) amino mercapto,phenyl, benzyl, alkoxyalkyl of up to 4 carbons or -(CH₂)_(n) COOH inwhich n is an integer of 1 to 4. Examples of suitable heterocyclicradicals include thienyl, furyl, pyrazolyl, imidazolyl, isoimidazolyl,triazolyl, tetrazolyl, thiazolyl, thiadiazolyl, oxazolyl, oxadiazolyl,isothiazolyl, isoxazolyl, pyridyl, pyridazinyl, pyrazinyl, pyrimidinyland triazinyl.

A most preferred group of compounds of formulae I and II are those inwhich Z is an optionally substituted 5- or 6- membered heterocyclic ringcontaining 1 to 4 atoms selected from N, O and S, the substituents beingpreferally those mentioned above. Especially preferred heterocyclicrings are optionally substituted triazole, thiadiazole, oxadiazole ortetrazole radicals. Within this group the most preferred Z substituentsare 1,2,3-triazolyl, 2-methyl-1,3,4-thiadiazol-5-yl,2-methyl-1,3,4-oxadiazol-5-yl, 1-N-methyltetrazolyl,1-carboxymethyltetrazol-5-yl and 1-carboxyethyltetrazol-5-yl.

The term "(lower)alkyl" as used herein means both staight and branchedchain aliphatic hydrocarbon radicals having from one to ten carbon atomssuch as methyl, ethyl, proyl, isopropyl, butyl, isobutyl, t-butyl, amyl,hexyl, 2-ethylhexyl, heptyl, decyl, etc. Similarly, where the term"(lower)" is used as part of the description of another group, e.g."(lower)alkoxy", it refers to the alkyl portion of such group which istherefore described above in connection with "(lower)alkyl".

The pharmaceutically acceptable salts referred to above include thenontoxic carboxylic acid salts, e.g. nontoxic metallic salts such assodium, potassium, calcium and aluminum, the ammonium salt and saltswith nontoxic amines, e.g. trialkylamines, procaine, dibenzylamine,N-benzyl-β-phenethylamine, 1-ephenamine, N,N'-dibenzylethylenediamine,N-alkylpiperidine and other amines which have been used to form salts ofpenicillins and cephalosporins. When a basic group is present, as whenit occurs in the 7-acyl group, the present invention also includes thepharmaceutically acceptable acid addition salts, e.g. salts with mineralacids such as hydrochloric, hydrobromic, hydroiodic, phosphoric,sulfuric and salts with organic acids such as maleic, acetic, citric,oxalic, succinic, benzoic, tartaric, fumaric, mandelic, ascorbic andmalic. The term "pharmaceutically acceptable salts" is also meant toinclude nontoxic acid addition salts of the easily cleavable estersreferred to above. The compounds which contain a basic group in radicalR may also be present in the form of an internal salt, i.e. in the formof the zwitterion.

The easily cleavable esters referred to above include ester groups whichare removable by methods, e.g. chemical or enzymatic hydrolysis,treatment with chemical reducing agents under mild conditions,irradiation with ultraviolet light or catalytic hydrogenation, which donot result in any appreciable destruction of the remaining portion ofthe molecule. Examples of suitable esters include those disclosed inU.S. Pat. Nos. 3,284,451 and 3,249,622 and U.K. Patents 1,229,453 and1,073,530. Esters which have been used previously in penicillin andcephalosporin chemistry include for example benzhydryl, p-nitrobenzyl,benzyl, trichloroethyl, silyl such as trimethylsilyl, phenacyl,p-methoxybenzyl, acetonyl, phthalidyl, indanyl and (lower)alkyl such asmethyl, ethyl and t-butyl. Particularly preferred easily cleavableesters are those which are hydrolyzed under physiological conditionssuch as pivaloyloxymethyl, acetoxymethyl, phthalidyl, indanyl andmethoxymethyl.

As the 0-2-isocephem compounds of the present invention may possess oneor more asymmetric carbon atoms, the invention includes all of thepossible enantiomeric and diastereomeric forms of the compounds of thegeneral formula II shown above. Resulting mixtures of isomers can beseparated into the individual isomers according to methods which areknown per se, e.g. fractional crystallization, adsorption chromatographyor other suitable separation processes. Resulting racemates can beseparated into the antipodes in the usual manner after introduction ofsuitable salt-forming groupings, e.g. by forming a mixture ofdiastereoisomeric salts with optically active salt-forming agents,separating the mixture into diastereoisomeric salts, and converting theseparated salts into the free compounds, or by fractionalcrystallization from optically active solvents.

It will be appreciated that certain of the compound of this inventionexist in various states of solvation and the anhydrous as well assolvated forms are within the scope of the invention.

The free acid compounds of general formula II and general formula Iwherein R is acylamido and physiologically hydrolyzed esters thereoftogether with the pharmaceutically acceptable salts of such free acidsand esters are useful as antibacterial agents. The remaining acids,esters and salts of formula I are valuable intermediates which can beconverted into the pharmacologically active compounds of formula II asby the processes described below.

Preferred compounds of formula II are those in which R is an acyl groupselected from the acyl groups defined above under (i) to (xvi). Use ofthe acyl groups mentioned above as being preferred within categories (i)to (xvi) results in active end-products having the most advantageouspharmacological properties.

A preferred embodiment of the present invention consists of thecompounds of formula II wherein R is an acyl group of the formula

i. R^(a) C_(n) H_(2n) Co-

wherein R^(a) is aryl, substituted aryl, cycloalkyl, substitutedcycloalkyl, cycloalkenyl, substituted cycloalkenyl or a nonaromatic ormesoionic heterocyclic group, and n is an integer from 1 to 4;

ii. C_(n) H_(2n-1) CO-

wherein n is an integer from 1 to 7, the alkyl portion of said acylgroup being straight or branched and optionally interrupted by an oxygenor sulphur atom;

iii. C_(n) H_(2n-1) CO-

wherein n is an integer from 2-7, the alkenyl portion of said acyl groupbeing straight or branched and optionally interrupted by an oxygen orsulfur atom; ##STR38## wherein R^(a) is as defined above under (i) andin addition may be benzyl, C₁ -C₆ alkyl or (lower)alkoxycarbonyl andR^(b) and R^(c) which may be the same or different each representhydrogen, phenyl, benzyl, phenethyl or C₁ -C₆ alkyl; ##STR39## whereinR^(a) is as defined above under (i) and in addition may be benzyl or C₁-C₆ alkyl and R^(b) and R^(c) are as defined under (iv);

vi. R^(a) X(CH₂)_(m) CO-

wherein R^(a) is as defined under (i) and in addition may be benzyl; Xis oxygen or sulfur; and m is an integer of 2-5;

vii. R^(a) CO--

wherein R^(a) is as defined under (i); ##STR40## wherein R^(a) is asdefined under (i) and Y is hydrazino, guanidino, ureido, thioureido,substituted thioureido, allophanamido, 3-guanyl-1-ureido, cyanamino,azido, 3-(2-furoyl)ureido, 3-(benzoyl)ureido, amino, acylamino, a groupobtained by reacting the amino group Y with an aldehyde or ketone,hydroxy, etherified hydroxy, esterified hydroxy, carboxy, esterifiedcarboxy, triazolyl, tetrazolyl, cyano, halogeno, acyloxy, sulfo,sulfoamino or esterified sulfo; ##STR41## wherein R^(d), R^(e) and R^(f)which may be the same or different may each represent C₁ -C₆ alkyl,phenyl or substituted phenyl; ##STR42## wherein R^(a) is as definedunder (i) and in addition may be hydrogen, C₁ -C₆ alkyl, halogen,substituted C₁ -C₆ alkyl, phenethyl, phenoxymethyl, benzyl or R^(a) -CO-and X is oxygen or sulfur; ##STR43## wherein Y is as defined under(viii) and n is an integer of 1-4;

xii. R^(g) CH(NH₂)-(CH₂)_(n) CO-

wherein n is an integer of 1-10 or

H₂ N-C_(n) H_(2n) Ar(CH₂)_(m) CO-

wherein m is 0 or an integer from 1-10 and n is 0, 1 or 2; R^(g) ishydrogen or an alkyl, aralkyl or carboxy group or a group as definedunder R^(a) in (i) above; and Ar is an arylene group;

xiii. R^(h) CO.sup.. CO-

wherein R^(h) is an aliphatic, araliphatic or aromatic group; ##STR44##wherein R^(a) is as defined under (i); X is oxygen or sulfur; X' isoxygen or imino and R^(i) is (lower)alkyl, C₄ -C₇ cycloalkyl, monohalo(lower)alkyl, dichloromethyl, trichloromethyl, (lower)alkenyl of 2-6carbon atoms, ##STR45## n is an integer from 0 to 3 inclusive and eachof R^(k) and R^(j) is hydrogen, nitro, di(lower)alkylamino,(lower)alkanoylamino, (lower)alkanoyloxy, C₁ -C₆ alkyl, C₁ -C₆ alkoxy,sulfamyl, chloro, iodo, bromo, fluoro or trifluoromethyl; ##STR46##wherein R^(a) is as defined under (i) and R^(i) is as defined under(xiv); or ##STR47## wherein R^(a) is as defined under (i) and R¹ is(lower)alkyl, C₃ -C₁₂ cycloalkyl, phenyl, a monocyclic heterocyclicradical having 5 or 6 atoms exclusive of hydrogen which are C, S, N orO, no more than 2 atoms being other than C, or a substituted monocyclicheterocyclic radical as defined above having one or more substituentsselected from halo, (lower)alkyl, (lower)alkoxy or phenyl.

Another preferred embodiment of the present invention consists of thecompounds of formula II wherein R is an acyl group of the formula

i. R^(a) C_(n) H_(2n) CO-

wherein R^(a) is (a) aryl selected from phenyl, 2-thienyl, 3-thienyl,furyl, 4-isoxazolyl, pyridyl, tetrazolyl, sydnone-3 or -4, imidazolyl,naphthoyl, quinoxalinyl, triazolyl, isothiazolyl, thiadiazolyl,thiazolyl, oxazolyl, oxadiazolyl, pyrazolyl, furazan, pyrazinyl,pyrimidinyl, pyridazinyl or triazinyl; (b) substituted aryl in which thearyl groups defined above under (a) are substituted by one or moreradicals selected from chloro, bromo, iodo, fluoro, nitro, amino, cyano,(lower)alkanoyloxy, (lower)alkanoyl, (lower)alkoxyamino, (lower)alkoxy,(lower)alkyl, (lower)alkylamino, hydroxy, guanidino, (lower)alkylthio,carboxy, phenyl, halophenyl, trifluoromethyl, di(lower)alkylamino,sulfamyl, (lower)alkanoylamino, phenyl(lower)alkylamido,cycloalkylamino, allylamido, morpholinocarbonyl, pyrrolidinocarbonyl,piperidinocarbonyl, tetrahydropyridino, furfurylamido orN-alkyl-N-anilino; (c) C₃ -C₁₂ cycloalkyl; (d) substituted C₃ -C₁₂cycloalkyl in which the substituents are one or more radicals selectedfrom chloro, bromo, fluoro, iodo, nitro, trifluoromethyl, C₁ -C₄ alkyl,C₁ -C₄ alkylamino, C₁ -C₂ alkoxy or amino; (e) C₃ -C₁₂ cycloalkenyl,said cycloalkenyl group having 1 or 2 double bonds; or (f) substitutedC₃ -C₁₂ cycloalkenyl, said cycloalkenyl group having 1 or 2 double bondsand being substituted by one or more radicals selected from chloro,bromo, fluoro, iodo, nitro, trifluoromethyl, C₁ -C₄ alkyl, C₁ -C₄alkylamino, C₁ -C₂ alkoxy or amino; and n is an integer from 1-4;

ii. C_(n) H_(2n) ⁺¹ CO-

wherein n is an integer from 1-7, the alkyl portion of said acyl groupbeing straight or branched and optionally interrupted by an oxygen orsulfur atom;

iii. C_(n) H_(2n-1) CO-

wherein n is an integer from 2-7, the alkenyl portion of said acyl groupbeing straight or branched and optionally interrupted by an oxygen orsulfur atom; ##STR48## wherein R^(a) is as defined above under (i) andin addition may be benzyl, C₁ -C₆ alkyl or (lower)alkoxy carbonyl andR^(b) and R^(c) which may be the same or different each representhydrogen, phenyl, benzyl, phenethyl or C₁ -C₆ alkyl; ##STR49## whereinR^(a) is as defined above under (i) and in addition may be benzyl or C₁-C₆ alkyl and R^(b) and R^(c) are as defined under (iv);

vi. R^(a) X(CH₂)_(m) CO-

wherein R^(a) is as defined under (i) and in addition may be benzyl; Xis oxygen or sulfur; and m is an integer of 2-5;

vii. R^(a) CO-

wherein R^(a) is as defined under (i); ##STR50## wherein R^(a) is asdefined under (i) and Y is hydrazino, guanidino, ureido; substitutedureido of the formula in which R^(p) is hydrogen or C₁ -C₈ alkyl andR^(q) is hydrogen, C₁ -C₈ alkyl, C₂ -C₈ alkenyl, phenyl, benzoyl, C₁ -C₈alkoxy-C₁ -C₈ alkyl or (carbo-C₁ -C₈ alkoxy)C₁ -C₈ alkyl; allophanamido;3-guanyl-1-ureido; 3-(2-furoyl)ureido; 3-(benzoyl)ureido; cyano;cyanamino; azido; amino; a group obtained by reacting the amino group Ywith acetone, formaldehyde, acetaldehyde, butyraldehyde, acetylacetone,methyl acetoacetate, benzaldehyde, salicylaldehyde, methyl ethyl ketoneor ethyl acetoacetate; hydroxy; (lower)alkoxy; carboxy;5-indanyloxycarbonyl; triazolyl; tetrazolyl; halogeno; formyloxy;(lower)alkanoyloxy; sulfo; or sulfoamino; ##STR51## wherein R^(d), R^(e)and R^(f) which may be the same or different may each represent C₁ -C₆alkyl, phenyl or phenyl substituted by one or more radicals selectedfrom chloro, bromo, iodo, fluoro, trifluoromethyl, nitro, amino, cyano,(lower)alkanoyloxy, (lower)alkanoyl, (lower)alkoxyamino, (lower)alkoxy,(lower)alkyl, (lower)alkylamino, hydroxy, (lower)alkylthio, carboxy,di(lower)alkylamino or sulfamyl; ##STR52## wherein R^(a) is as definedunder (i) and in addition may be hydrogen, C₁ -C₆ alkyl,halogen-substituted C₁ -C₆ alkyl, phenethyl, phenoxymethyl, benzyl orR^(a) -CO- and X is oxygen or sulfur. ##STR53## wherein Y is as definedunder (viii) and n is an integer of 1-4;

xii. R^(g) CH(NH₂)(CH₂)_(n) CO-

wherein n is an integer of 1-10, or

H₂ N-C_(n) H_(2n) Ar(CH₂)_(m) CO-

wherein m is 0 or an integer from 1-10, and n is 0, 1 or 2; R^(g) ishydrogen, (lower)alkyl, phenyl, benzyl or carboxy and Ar is p-phenyleneor 1,4naphthylene;

xiii. R^(h) Co.Co-

wherein R^(h) is 2-thienyl; 3-thienyl; α-naphthyl; 2-phenanthryl or amono-, di- or tri-substituted phenyl group, the substituents beingselected from chloro, bromo, iodo, fluoro, amino, di(lower)alkylamino,(lower)alkyl, (lower)alkoxy, nitro or (lower)alkanoylamino; ##STR54##wherein R^(a) is as defined under (i); X is oxygen or sulfur; X' isoxygen or imino; and R^(i) is (lower)alkyl, cycloalkyl having 4, 5, 6 or7 carbon atoms, monohalo (lower)alkyl, dichloromethyl, trichloromethyl,(lower)alkenyl of 2-6 carbon atoms, ##STR55## n is an integer from 0 to3 inclusive and each of R^(k) and R^(j) is hydrogen, nitro,di(lower)alkylamino, (lower)alkanoylamino, (lower)-alkanoyloxy, C₁ -C₆alkyl, C₁ -C₆ alkoxy, sulfamyl, chloro, iodo, bromo, fluoro, ortrifluoromethyl; wherein R^(a) is as defined under (i) and R^(i) is asdefined under (xiv); or ##STR56## wherein R^(a) is as defined under (i)and R¹ is (lower)alkyl, cycloalkyl of 3-12 carbon atoms, phenyl, amonocyclic heterocyclic radical having 5 or 6 atoms exclusive ofhydrogen which are C, S, N or O, no more than 2 atoms being other thanC, or a substituted monocyclic heterocyclic radical as defined abovehaving one or more substituents selected from halo, (lower)alkyl,(lower)alkoxy or phenyl.

Another preferred embodiment of the present invention consists of thecompounds of formula II wherein R is as defined immediately above and Zis C₁ -C₆ alkyl, aryl selected from phenyl or naphthyl, aralkyl of theformula aryl -- (ALK)_(m) -- in which aryl is phenyl or naphthyl, m isan integer of 1 to 4 and ALK represents a straight or branched chainalkylene radical, or heterocyclic selected from a 5- or 6- memberedheterocyclic ring containing 1 to 4 atoms selected from N, O or S, saidalkyl radical being optionally substituted by one or more substituentsselected from hydroxy, halo, amino, nitro, di (C₁ -C₄ alkyl) amino,carboxy, sulfo or cyano and said aryl, aralkyl or heterocyclic radicalsbeing optionally substituted by one or more substituents selected fromhalo, C₁ -C₄ alkyl, C₁ -C₄ alkoxy, cyano, carboxyl, amino, nitro, C₃ -C₄cycloalkyl, C₂ -C₄ alkenyl, trifluoromethyl, hydroxy, hydroxymethyl, C₁-C₄ alkylthio, C₁ -C₄ alkylamino, di (C₁ -C₄ alkyl)amino, mercapto,phenyl, benzyl, alkoxyalkyl of up to 4 carbons or -(CH₂)_(n) COOH inwhich n is an integer of 1 to 4.

Within this group of compounds a preferred subclass consists of thecompounds wherein R is an acyl group of the formula ##STR57## whereinAr' is a radical of the formula ##STR58## in which R^(m), R^(n) andR^(o) are alike or different and each is hydrogen, hydroxy,(lower)alkyl, cyano, (lower)alkoxy, chloro, bromo, iodo, fluoro,trifluoromethyl, nitro, amino, (lower)alkylamino, di(lower)alkylamino,(lower)alkanoyl, (lower)alkanoyloxy or phenyl and Y is amino or a groupobtained by reacting the amino group with acetaldehyde, formaldehyde oracetone; fluoro; chloro; bromo; iodo; hydroxy; (lower)alkanoyloxy;carboxy; guanidino; 3-guanyl-1-ureido; 3-(2-furoyl)ureido;3-benzoylureido; sulfo; sulfoamino; ureido; thioureido; (lower)alkoxy;cyano; cyanamino; or indanyloxycarbonyl.

A second preferred subclass within this group of compounds consists ofthe compounds wherein R is an acyl group of the formula ##STR59##

Another preferred embodiment of the present invention consists of thecompounds of formula ##STR60## in which R is an acyl group and Zrepresents a 5- or 6-membered heterocyclic ring containing N, O or S,said heterocyclic ring being optionally substituted by one or moresubstituents selected from halo, C₁ -C₄ alkyl, C₁ -C₄ alkoxy, cyano,carboxyl, amino, nitro, C₃ -C₄ cycloalkyl, C₂ -C₄ alkenyl,trifluoromethyl, hydroxy, hydroxymethyl, C₁ -C₄ alkylthio, C₁ -C₄alkylamino, di (C₁ -C₄ alkyl) amino, mercapto, phenyl, benzyl,alkoxyalkyl of up to 4 carbons or --(CH₂)_(n) COOH in which n is aninteger of 1 to 4, and easily cleavable esters and pharmaceuticallyacceptable salts thereof.

Within this group of compounds, a preferred subclass consists of thecompounds wherein R is an acyl group of the formula

i. R^(a) C_(n) H_(2n) CO--

wherein R^(a) is aryl, substituted aryl, cycloalkyl, substitutedcycloalkyl, cycloalkenyl, substituted cycloalkenyl or a nonaromatic ormesoionic heterocyclic group, and n is an integer from 1 to 4;

ii. C_(n) H_(2n-1) CO--

wherein n is an integer from 1-7, the alkyl portion of said acyl groupbeing straight or branched and optionally interrupted by an oxygen orsulphur atom; iii. C_(n) H_(2n-1) CO--

wherein n is an integer from 2-7, the alkenyl portion of said acyl groupbeing straight or branched and optionally interrupted by an oxygen orsulfur atom; ##STR61## wherein R^(a) is as defined above under (i) andin addition may be benzyl, C₁ -C₆ alkyl or (lower)alkoxycarbonyl andR^(b) and R^(c) which may be the same or different each representhydrogen, phenyl, benzyl, phenethyl or C₁ -C₆ alkyl; ##STR62## whereinR^(a) is as defined above under (i) and in addition may be benzyl or C₁-C₆ alkyl and R^(b) and R^(c) are as defined under (iv);

vi. R^(a) X(CH₂)_(m) CO--

wherein R^(a) is as defined under (i) and in addition may be benzyl; Xis oxygen or sulfur; and m is an integer of 2-5; vii. R^(a) CO--

wherein R^(a) is as defined under (i); ##STR63## wherein R^(a) is asdefined under (i) and Y is hydrazino, guanidino, ureido, thioureido,substituted thioureido, allophanamido, 3-guanyl-1-ureido, cyanamino,azido, 3-(2-furoyl)ureido, 3-(benzoyl)ureido, amino, acylamino, a groupobtained by reacting the amino group Y with an aldehyde or ketone,hydroxy, etherified hydroxy, esterified hydroxy, carboxy, esterifiedcarboxy, triazolyl, tetrazolyl, cyano, halogeno, acyloxy, sulfo,sulfoamino or esterified sulfo; ##STR64## wherein R^(d), R^(e) and R^(f)which may be the same or different may each represent C₁ -C₆ alkyl,phenyl or substituted phenyl; ##STR65## wherein R^(a) is as definedunder (i) and in addition may be hydrogen, C₁ -C₆ alkyl, halogen,substituted C₁ -C₆ alkyl, phenethyl, phenoxymethyl, benzyl or R^(a) -CO-and X is oxygen or sulfur; ##STR66## wherein Y is as defined under(viii) and n is an integer of 1-4;

xii. R^(g) CH(NH₂)--(CH₂)_(n) CO--

wherein n is an integer of 1-10 or

H₂ N-C_(n) H_(2n) Ar(CH₂)_(m) CO--

wherein m is 0 or an integer from 1-10 and n is 0, 1 or 2; R^(g) ishydrogen or an alkyl, aralkyl or carboxy group or a group as definedunder R^(a) in (i) above; and Ar is an arylene group;

xiii. R^(h) CO.CO--

wherein R^(h) is an aliphatic, araliphatic or aromatic group; ##STR67##wherein R^(a) is as defined under (i); X is oxygen or sulfur; X' isoxygen or imino and R^(i) is (lower)alkyl, C₄ -C₇ cycloalkyl, monohalo(lower)alkyl, dichloromethyl, trichloromethyl, (lower)alkenyl of 2-6carbon atoms, ##STR68## n is an integer from 0 to 3 inclusive and eachof R^(k) and R^(j) is hydrogen, nitro, di(lower)alkylamino,(lower)alkanoylamino, (lower)-alkanoyloxy, C₁ -C₆ alkyl, C₁ -C₆ alkoxy,sulfamyl, chloro, iodo, bromo, fluoro or trifluoromethyl; ##STR69##wherein R^(a) is as defined under (i) and R^(i) is as defined under(xiv); or wherein R^(a) is as defined under (i) and R¹ is (lower)alkyl,C₃ -C₁₂ cycloalkyl, phenyl, a monocyclic heterocyclic radical having 5or 6 atoms exclusive of hydrogen which are C, S, N or O, no more than 2atoms being other than C, or a substituted monocyclic heterocyclicradical as defined above having one or more substituents selected fromhalo, (lower)alkyl, (lower)alkoxy or phenyl.

Another preferred subclass within this group consists of the compoundswherein R is an acyl group of the formula

i. R^(a) C_(n) H_(2n) CO--

wherein R^(a) is (a) aryl selected from phenyl, 2-thienyl, 3-thienyl,furyl, 4-isoxazolyl, pyridyl, tetrazoyl, sydnone-3 or -4, imidazolyl,naphthoyl, quinoxalinyl, triazolyl, isothiazolyl, thiadiazolyl,thiazolyl, oxazolyl, oxadiazolyl, pyrazolyl, furazan, pyrazinyl,pyrimidinyl, pyridazinyl or triazinyl; (b) substituted aryl in which thearyl groups defined above under (a) are substituted by one or moreradicals selected from chloro, bromo, iodo, fluoro, nitro, amino, cyano,(lower)alkanoyloxy, (lower)alkanoyl, (lower)alkoxyamino, (lower)alkoxy,(lower)alkyl, (lower)alkylamino, hydroxy, guanidino, (lower)alkylthio,carboxy, phenyl, halophenyl, trifluoromethyl, di(lower)alkylamino,sulfamyl, (lower)alkanoylamino, phenyl(lower)alkylamido cycloalkylamino,allylamido, morpholinocarbonyl, pyrrolidinocarbonyl, piperidinocarbonyl,tetrahydropyridino, furfurylamido or N-alkyl-N-anilino; (c) C₃ -C₁₂cycloalkyl; (d) substituted C₃ -C₁₂ cycloalkyl in which the substituentsare one or more radicals selected from chloro, bromo, fluoro, iodo,nitro, trifluoromethyl, C₁ -C₄ alkyl, C₁ -C₄ alkylamino, C₁ -C₂ alkoxyor amino; (e) C₃ -C₁₂ cycloalkenyl, said cycloalkenyl group having 1 or2 double bonds; or (f) substituted C₃ -C₁₂ cycloalkenyl, saidcycloalkenyl group having 1 or 2 double bonds and being substituted byone or more radicals selected from chloro, bromo, fluoro, iodo, nitro,trifluoromethyl, C₁ -C₄ alkyl, C₁ -C₄ alkylamino, C₁ -C₂ alkoxy oramino; and n is an integer from 1-4;

ii. C_(n) H_(2n) ₊₁ CO--

wherein n is an integer from 1-7, the alkyl portion of said acyl groupbeing straight or branched and optionally interrupted by an oxygen orsulfur atom;

iii. C_(n) H_(2n-1) CO--

wherein n is an integer from 2-7, the alkenyl portion of said acyl groupbeing straight or branched and optionally interrupted by an oxygen orsulfur atom; ##STR70## wherein R^(a) is as defined above under (i) andin addition may be benzyl, C₁ -C₆ alkyl or (lower)alkoxycarbonyl andR^(b) and R^(c) which may be the same or different each representhydrogen, phenyl, benzyl, phenethyl or C₁ -C₆ alkyl; ##STR71## whereinR^(a) is as defined above under (i) and in addition may be benzyl or C₁-C₆ alkyl and R^(b) and R^(c) are as defined under (iv);

vi. R^(a) X(CH₂) mCO--

wherein R^(a) is as defined under (i) and in addition may be benzyl; Xis oxygen or sulfur; and m is an integer of 2-5;

vii. R^(a) CO--

wherein R^(a) is as defined under (i); ##STR72## wherein R^(a) is asdefined under (i) and Y is hydrazino, guanidino, ureido; substitutedureido of the formula ##STR73## in which R^(p) is hydrogen or C₁ -C₈ andR^(q) is hydrogen, C₁ -C₈ alkyl, C₂ -C₈ alkenyl, phenyl, benzoyl, C₁ -C₈alkoxy-C₁ -C₈ alkyl or (carbo-C₁ -C₈ alkoxy)C₁ -C₈ alkyl; allophanamido;3-guanyl-1-ureido; 3-(2-furoyl)ureido; 3-(benzoyl)ureido; cyano;cyanamino; azido; amino; a group obtained by reacting the amino group Ywith acetone, formaldehyde, acetaldehyde, butyraldehyde, acetylacetone,methyl acetoacetate, benzaldehyde, salicylaldehyde, methyl ethyl ketoneor ethyl acetoacetate; hydroxy; (lower)alkoxy; carboxy;5-indanyloxycarbonyl; triazolyl; tetrazolyl; halogeno; formyloxy;(lower)alkanoyloxy; sulfo; or sulfoamino; ##STR74## wherein R^(d), R^(e)and R^(f) which may be the same or different may each represent C₁ -C₆alkyl, phenyl or phenyl substituted by one or more radicals selectedfrom chloro, bromo, iodo, fluoro, trifluoromethyl, nitro, amino, cyano,(lower)alkanoyloxy, (lower)alkanoyl, (lower)alkoxyamino, (lower)alkoxy,(lower)alkyl, (lower)alkylamino, hydroxy, (lower)alkylthio, carboxy,di(lower)alkylamino or sulfamyl; ##STR75## wherein R^(a) is as definedunder (i) and in addition may be hydrogen, C₁ -C₆ alkyl, halogen-substituted C₁ -C₆ alkyl, phenethyl, phenoxymethyl, benzyl or R^(a)--CO-- and X is oxygen or sulfur. ##STR76## wherein Y is as definedunder (viii) and n is an integer of 1-4;

xii. R^(g) CH(NH₂) (CH₂)_(n) CO--

wherein n is an integer of 1-10, or

H₂ H-C_(n) H_(2n) Ar(CH₂)_(m) CO--

wherein m is 0 or an integer from 1-10, and n is 0, 1 or 2; R^(g) ishydrogen, (lower)alkyl, phenyl, benzyl or carboxy and Ar is p-phenyleneor 1,4-naphthylene;

xiii. R^(h) CO CO--

wherein R^(h) is 2-thienyl; 3-thienyl; α-naphthyl; 2-phenanthryl or amono-, di- or tri-substituted phenyl group, the substituents beingselected from chloro, bromo, iodo, fluoro, amino, di(lower)alkylamino,(lower)alkyl, (lower)alkoxy, nitro or (lower)alkanoylamino; ##STR77##wherein R^(a) is as defined under (i); X is oxygen or sulfur; X' isoxygen or imino; and R^(i) is (lower)alkyl, cycloalkyl having 4, 5, 6 or7 carbon atoms, monohalo (lower)alkyl, dichloromethyl, trichloromethyl,(lower)alkenyl of 2-6 carbon atoms, ##STR78## n is an integer from 0 to3 inclusive and each of R^(k) and R^(j) is hydrogen, nitro,di(lower)-alkylamino, (lower)alkanoylamino, (lower)alkanoyloxy, C₁ -C₆alkyl, C₁ -C₆ alkoxy, sulfamyl, chloro, iodo, bromo, fluoro, ortrifluoromethyl; wherein R^(a) is as defined under (i) and R^(i) is asdefined under (xiv); or ##STR79## wherein R^(a) is as defined under (i)and R¹ is (lower)alkyl, cycloalkyl of 3-12 carbon atoms, phenyl, amonocyclic heterocyclic radical having 5 or 6 atoms exclusive ofhydrogen which are C, S, N or O, no more than 2 atoms being other thanC, or a substituted monocyclic heterocyclic radical as defined abovehaving one or more substituents selected from halo, (lower)alkyl,(lower)alkoxy or phenyl.

Another preferred subclass within this group consists of the compoundswherein R is an acyl group of the formula

R^(a) C_(n) H_(2n) CO--

in which R^(a) is phenyl; phenyl substituted by one or more radicalsselected from chloro, bromo, iodo, fluoro, nitro, amino, (lower) alkyl,guanidino, (lower) alkylthio, cyano, (lower) alkoxy, sulfamyl, (lower)alkylamino, hydroxy, acetoxy or trifluoromethyl; 2-thienyl; 3-thienyl;tetrazolyl; sydnone-3- or sydnone-4; furyl; isothiazolyl; thiadiazolyloptionally substituted with phenyl; oxadiazolyl optionally substitutedwith phenyl; thiazolyl; imidazolyl; triazolyl; oxazolyl, pyridyl;furazan optionally substituted at the 3-position with methoxy;4-isoxazolyl optionally substituted at the 5-position with methyl and atthe 3-position with phenyl or halophenyl; 1, 4-cyclohexadienyl;1-cyclohexenyl and 1-aminocyclohexyl. The most preferred compounds ofthis subclass are those in which n is 1.

Another preferred subclass within this group consists of the compoundswherein R is an acyl group of the formula ##STR80## in which R^(a) isphenyl and R^(b) and R^(c) which may be the same or different eachrepresent hydrogen, phenyl, benzyl, phenethyl or C₁ -C₆ alkyl.

Another preferred subclass within this group consists of the compoundswherein R is an acyl group of the formula ##STR81## in which R^(a) isphenyl; phenyl substituted with one or more radicals selected fromchloro, bromo, iodo, fluoro, nitro, amino, (lower)alkyl,(lower)alkylthio, cyano, (lower)alkoxy, (lower)alkylamino, hydroxy,acetoxy or trifluoromethyl; 3-pyridyl or 4-pyridyl; and R^(b) and R^(c)are hydrogen.

Another preferred subclass within this group consists of the compoundswherein R is an acyl group of the formula R^(a) CO-- in which R^(a) isphenyl; phenyl substituted with one or more radicals selected fromchloro, bromo, iodo, fluoro, nitro, amino, (lower)alkyl,(lower)alkylthio, cyano, (lower)alkoxy, (lower)alkylamino,di(lower)alkylamino, hydroxy, acetoxy or trifluoromethyl;2-ethoxynaphthoyl; 3-phenyl-5-methylisoxazol-4-yl;3-o-chlorophenyl-5-methylisoxazol-4-yl;3-(2,6-dichlorophenyl)-5-methylisoxazol-4yl; or 1-aminocyclohexyl.

Another preferred subclass within this group consists of the compoundswherein R is an acyl group of the formula ##STR82## in which R^(a) is(a) aryl selected from phenyl, 2-thienyl, 3-thienyl, furyl,4-isoxazolyl, pyridyl, tetrazolyl, sydnone-3 or -4, imidazolyl,naphthoyl, quinoxalinyl, triazolyl, isothiazolyl, thiadiazolyl,thiazolyl, oxazolyl, oxadiazolyl, pyrazolyl, furazan, pyrazinyl,pyrimidinyl, pyridazinyl or triazinyl; (b) substituted aryl in which thearyl groups defined above under (a) are substituted by one or moreradicals selected from chloro, bromo, iodo, fluoro, nitro, amino, cyano,(lower)alkanoyloxy, (lower)alkanoyl, (lower)alkoxyamino, (lower)alkoxy,(lower)alkyl, (lower)alkylamino, hydroxy, guanidino, (lower)alkythio,carboxy, phenyl, halophenyl, trifluoromethyl, di(lower)alkylamino,sulfamyl, (lower)alkanoylamino, phenyl(lower)alkylamido,cycloalkylamino, allylamido, morpholinocarbonyl, pyrrolidinocarbonyl,piperidinocarbonyl, tetrahydropyridino, furfurylamido orN-alkyl-N-anilino; (c) C₃ -C₁₂ cycloalkyl; (d) substituted C₃ -C₁₂cycloalkyl in which the substituents are one or more radicals selectedfrom chloro, bromo, fluoro, iodo, nitro, trifluoromethyl, C₁ -C₄ alkyl,C₁ -C₄ alkylamino, C₁ -C₂ alkoxy or amino; (e) C₃ -C₁₂ cycloalkenyl,said cycloalkenyl group having 1 or 2 double bonds; or (f) substitutedC₃ -C₁₂ cycloalkenyl, said cycloalkenyl group having 1 or 2 double bondsand being substituted by one or more radicals selected from chloro,bromo, fluoro, iodo, nitro, trifluoromethyl, C₁ -C₄ alkyl, C₁ -C₄alkylamino, C₁ -C₂ alkoxy or amino; and Y is hydrazino, guanidino,ureido; substituted ureido of the formula ##STR83## in which R^(p) ishydrogen or C₁ -C₈ alkyl and R^(q) is hydrogen, C₁ -C₈ alkyl, C₂ -C₈alkenyl, phenyl, benzoyl, C₁ -C₈ alkoxy-C₁ -C₈ alkyl or (carbo--C₁ -C₈alkoxy)C₁ -C₈ alkyl; allophanamido; 3-guanyl-1-ureido;3-(2-furoyl)ureido; 3-(benzoyl)ureido; cyano; cyanamino; azido; amino; agroup obtained by reacting the amino group Y with acetone, formaldehyde,acetaldehyde, bu yraldehyde, acetylacetone, methyl acetoacetate,benzaldehyde, salicylaldehyde, methyl ethyl ketone or ethylacetoacetate; hydroxy; (lower)alkoxy; carboxy; 5-indanyloxycarbonyl;triazolyl; tetrazolyl; halogeno; formyloxy; (lower)alkanoyloxy; sulfo;or sulfoamino;

Another preferred subclass within this group consists of the compoundswherein R is an acyl group of the formula ##STR84## in which R^(a) is2-thienyl; 3-thienyl; phenyl; or phenyl substituted by one or moreradicals selected from nitro, di(lower)alkylamino, (lower)alkanoylamino,amino, hydroxy, (lower)alkanoyloxy, C₁ -C₆ alkyl, C₁ -C₆ alkoxy,sulfamyl, chloro, bromo, iodo, fluoro, or trifluoromethyl; X is oxygen;X' is oxygen or imino; and R^(i) is (lower)alkyl, phenyl, 2-thienyl,3-thienyl, 2-furyl or 5-nitro-2-furyl. The most preferred compounds ofthis subclass are those in which R^(a) is phenyl, p-hydroxyphenyl,2-thienyl or 3-thienyl, X' is oxygen and R^(i) is phenyl or 2-furyl.

A most preferred subclass within this group consists of the compoundswherein R is an acyl group of the formula ##STR85## in which Ar' is aradical of the formula ##STR86## in which R^(m), R^(n) and R^(o) arealike or different and each is hydrogen, hydroxy, (lower)alkyl, cyano,(lower)alkoxy, chloro, bromo, iodo, fluoro, trifluoromethyl, nitro,amino, (lower)alkylamino, di(lower)alkylamino, (lower)alkanoyl,(lower)alkanoyloxy or phenyl and Y is amino or a group obtained byreacting the amino group with acetaldehyde, formaldehyde or acetone;fluoro; chloro; bromo; iodo; hydroxy; (lower)alkanoyloxy; carboxy;guanidino; 3-guanyl-1-ureido; 3-(2-furoyl)ureido; 3-benzoylureido;sulfo; sulfoamino; ureido; thioureido; (lower)alkoxy; cyano; cyanamino;or indanyloxycarbonyl. The most preferred compounds of this subclass arethose in which Ar' is phenyl, p-hydroxyphenyl, 4-hydroxy-3,5-dichlorophenyl, 3-chloro-4-hydroxyphenyl, o-, m- orp-aminomethylphenyl, 2-thienyl, 3-thienyl, 1-cyclohexenyl or 1,4-cyclohexadienyl and Y is amino, hydroxy or carboxy.

A most preferred subclass within this group consists of the compoundswherein R is an acyl group of the formula ##STR87##

A most preferred subclass within this group consists of the compoundswherein R is an acyl group of the formula ##STR88##

A most preferred subclass within this group consists of the acids inwhich R is a-carboxyphenylacetyl, cyanoacetyl,α-amino-α-(p-hydroxyphenyl)acetyl,α-amino-α-(3-chloro-4-hydroxyphenyl)acetyl,α-amino-α-(3,5-dichloro-4-hydroxyphenyl)acetyl,α-amino-α-(2-thienyl)acetyl, α-amino-α-(3-thienyl)acetyl,α-amino-α-(1-cyclohexenyl)acetyl, α-amino-α-(1,4-cyclohexadienyl)acetyl,α-hydroxyacetyl, α-hydroxy-α-(2-thienyl)acetyl,α-hydroxy-α-(3-thienyl)acetyl, α-hydroxy-α-(1-cyclohexenyl)acetyl,α-hydroxy-α-(1,4-cyclohexadienyl)acetyl, α-carboxy-α-(2-thienyl)acetyl,α-carboxy-α-(3-thienyl)acetyl, α-carboxy-α-(1-cyclohexenyl)acetyl,α-carboxy-α-(1,4-cyclohexadienyl)acetyl,α-indanyloxycarbonyl-α-phenylacetyl, 1-(1H)tetrazolyl,4-pyridylthioacetyl, 2-thienylacetyl, 3-thienylacetyl,1-cyclohexenylacetyl, 1,4-cyclohexadienylacetyl,α-aminomethylphenylacetyl, 1-aminocyclohexylcarbonyl,2,6-dimethoxybenzoyl, sydnoneacetyl or α-azidophenylacetyl, andpharmaceutically acceptable salts thereof.

A most preferred subclass within this group consists of the D-isomers ofthe acids in which R is α-amino-α-(p-hydroxyphenyl)acetyl,α-amino-α-(3-chloro-4-hydroxyphenyl)acetyl,α-amino-α-(3,5-dichloro-4-hydroxyphenyl)acetyl,α-amino-α-(2-thienyl)acetyl, α-amino-α-(3-thienyl)acetyl,α-amino-α-(1-cyclohexenyl)acetyl, α-amino-α-(1,4-cyclohexadienyl)acetyl,α-hydroxyacetyl, α-hydroxy-α-(2-thienyl)acetyl,α-hydroxy-α-(3-thienyl)acetyl, α-hydroxy-α-(1-cyclohexenyl)acetyl or aα-hydroxy-α-(1,4-cyclohexadienyl)acetyl, or pharmaceutically acceptablesalts thereof.

Another preferred embodiment of the present invention consists of thecompounds of formula ##STR89## wherein R is an acyl group and Z is atriazole, thiadiazole, oxadiazole or tetrazole radical, said radicalbeing optionally substituted by one or more substituents selected fromhalo, C₁ -C₄ alkyl, C₁ -C₄ alkoxy, cyano, carboxyl, amino, nitro, C₃ -C₄cycloalkyl, C₂ -C₄ alkenyl, trifluoromethyl, hydroxy, hydroxymethyl, C₁-C₄ alkylthio, C₁ -C₄ alkylamino, di (C₁ -C₄ alkyl)amino, phenyl,benzyl, mercapto, alkoxyalkyl of up to 4 carbons or --(CH₂)_(n) COOH inwhich n is an integer of 1 to 4, and easily cleavable esters andpharmaceutically acceptable salts thereof.

A most preferred embodiment of the present invention consists of thecompounds of formula II in which Z is 1,2,3-triazol-5-yl,2-methyl-1,3,4-thiadiazol-5-yl, 2-methyl-1,3,4-oxadiazol-5-yl,1-N-methyltetrazol-5-yl, 1-carboxymethyltetrazol-5-yl or1-carboxyethyltetrazol-5-yl.

A preferred subclass within this group consists of the compounds inwhich R is ##STR90## in which Ar' is a radical of the formula ##STR91##in which R^(m), R^(n) and R^(o) are alike or different and each ishydrogen, hydroxy, (lower)alkyl, cyano, (lower)alkoxy, chloro, bromo,iodo, fluoro, trifluoromethyl, nitro, amino, (lower)alkylamino,di(lower)alkylamino, (lower)alkanoyl, (lower)alkanoyloxy or phenyl and Yis amino or a group obtained by reacting the amino group withacetaldehyde, formaldehyde or acetone; fluoro; chloro; bromo; iodo;hydroxy; (lower)alkanoyloxy; carboxy; guanidino; 3-guanyl-1-ureido;3-(2-furoyl)ureido; 3-benzoylureido; sulfo; sulfoamino; ureido;thioureido; (lower)alkoxy; cyano; cyanamino; or indanyloxycarbonyl.

The most preferred compounds of this subclass are those in which Ar' isphenyl, p-hydroxyphenyl, 4-hydroxy-3,5-dichlorophenyl,3-chloro-4-hydroxyphenyl, o-, m- or p-aminomethylphenyl, 2-thienyl,3-thienyl, 1-cyclohexenyl or 1,4-cyclohexadienyl and Y is amino, hydroxyor carboxy.

Another preferred subclass within this group consists of the compoundswherein R is an acyl group of the formula ##STR92## Another preferredsubclass within this group consists of the compounds in which R is anacyl group of the formula ##STR93## wherein U and V are alike ordifferent and each is hydrogen, chloro or fluoro ##STR94##

Another preferred subclass within this group consists of the acids inwhich R is α-carboxyphenylacetyl, cyanoacetyl,α-amino-α-(p-hydroxyphenyl)acetyl,α-amino-α-(3-chloro-4-hydroxyphenyl)acetyl,α-amino-α-(3,5-dichloro-4-hydroxyphenyl)acetyl,α-amino-α-(2-thienyl)acetyl, α-amino-α-(3-thienyl)acetyl,α-amino-α-(1-cyclohexenyl)acetyl, α-amino-α-(1,4-cyclohexadienyl)acetyl,α-hydroxyacetyl, α-hydroxy-α-(2-thienyl)acetyl,α-hydroxy-α-(3-thienyl)acetyl, α-hydroxy-α-(1-cyclohexenyl)acetyl,α-hydroxy-α-)1,4-cyclohexadienyl)acetyl, α-carboxy-α-(2-thienyl)acetyl,α-carboxy-α-(3-thienyl)acetyl, α-carboxy-α-(1-cyclohexenyl)acetyl,α-carboxy-α-(1,4-cyclohexadienyl)acetyl,α-indanyloxycarbonyl-α-phenylacetyl, 1-(1H)tetrazolyl,4-pyridylthioacetyl, 2-thienylacetyl, 3-thienylacetyl,1-cyclohexenylacetyl, 1,4-cyclohexadienylacetyl,o-aminomethylphenylacetyl, 1-aminocyclohexylcarbonyl,2,6-dimethoxybenzoyl, sydnoneacetyl or α-azidophenylacetyl, orpharmaceutically acceptable salts thereof.

Another preferred subclass within this group consists of the D-isomersof the acids in which R is α-amino-α-(p-hydroxyphenyl)acetyl,α-amino-α-(3-chloro-4-hydroxyphenyl)acetyl,α-amino-α-(3,5-dichloro-4-hydroxyphenyl)acetyl,α-amino-α-(2-thienyl)acetyl, α-amino-α-(3-thienyl)acetyl,α-amino-α-(1-cyclohexenyl)acetyl, α-amino-α-(1,4-cyclohexadienyl)acetyl,α-hydroxyacetyl, α-amino-α-(2-thienyl)acetyl,α-hydroxy-α-(3-thienyl)acetyl, α-hydroxy-α-(1-cyclohexenyl)acetyl orα-hydroxy-α-(1,4-cyclohexadienyl)acetyl, or pharmaceutically acceptablesalts thereof.

The present invention further provides various novel intermediatesuseful in the synthesis of the 7-acylamido 0-2-isocephem compounds offormula II described above.

Preferred embodiments of the present invention are the novelintermediates having the formula ##STR95## wherein Z is optionallysubstituted C₁ -C₆ alkyl, aryl, aralkyl or heterocyclic and R" ishydrogen or an easily cleavable ester carboxyl-protecting group, andsalts thereof. The preferred Z substituents are as defined above inconnection with the compounds of formulae I and II.

Other preferred embodiments of the present invention are theintermediates having the formula ##STR96## wherein Z is optionallysubstituted C₁ -C₆ alkyl, aryl, aralkyl or heterocyclic and R" ishydrogen or an easily cleavable ester carboxyl-protecting group, andsalts thereof. The preferred Z substituents are those mentioned above asbeing preferred in connection with the compounds of formulae I and II.

The intermediates of formulae III and IV may be in the form of the freecarboxylic acid or a salt thereof or in the form where the carboxylgroup is protected in a conventional manner such as preferably byesterification. The protecting group is selected so that it may beremoved by methods which do not result in any appreciable destruction ofthe remaining portion of the molecule. Preferred carboxyl protectinggroups are the easily cleavable esters as defined above including inparticular benzhydryl, p-nitrobenzyl, trichloroethyl, silyl includingespecially trimethylsilyl, phenacyl, p-methoxybenzyl, acetonyl,(lower)alkyl such as methyl, t-butyl or ethyl, benzyl, triphenylmethyl,methoxymethyl, acetoxymethyl, phthalidyl, indanyl and pivaloyloxymethyl.

The novel 7-acylamido compounds of the formula II may be prepared byN-acylating a 7-amino intermediate of the formula ##STR97## wherein Z isoptionally substituted C₁ -C₆ alkyl, aryl, aralkyl or heterocyclic andR" is hydrogen or an easily cleavable ester carboxyl-protecting group,or a salt thereof, with an acylating acid of the formula

R-COOH

wherein R is an acyl group, or with its functional equivalent as anacylating agent for a primary amine and, if desired, (a) when R" is acarboxyl-protecting group, converting the 7-acylated ester to the freeacid compound or a physiologically hydrolyzed ester or apharmaceutically acceptable salt of said acid or ester, or (b) when R"is hydrogen, converting the 7-acylated carboxylic acid to aphysiologically hydrolyzed ester or a pharmaceutically acceptable saltof said acid or ester and, if desired, resolving a resulting isomermixture into its component isomers.

The 7-amino starting materials of general formula III are of useprimarily as intermediates in preparing the pharmacologically activeN-acyl derivatives of formula II. The free acids, physiologicallyhydrolyzed esters and pharmaceutically acceptable salts of said acidsand esters of formula III, however, do possess some antibacterialactivity per se against various pathogenic microorganisms.

The 7-acylamido 0-2-isocephem compounds of formula II are prepared byN-acylation according to known methods of the 7-amino group ofintermediate III with an acylating acid of the formula

R-COOH

wherein R is an acyl group, or with its functional equivalent as anacylating agent for a primary amino group. The acylating agents forpreparing the products of formula II are known, readily preparable byknown methods or described herein.

Intermediate III may be acylated either in the form of the freecarboxylic acid (or salt thereof) or as an easily cleavable ester (oracid addition salt thereof). Preferred esters include benzhydryl,benzyl, p-nitrobenzyl, trichloroethyl, silyl (especiallytrimethylsilyl), phenacyl, p-methoxybenzyl, acetonyl, (lower)alkylincluding particularly methyl, ethyl and t-butyl, triphenylmethyl,methoxymethyl, acetoxymethyl, pivaloyloxymethyl, phthalidyl and indanyl.The procedures for preparing esters of carboxylic acids are disclosed inthe literature and are well-known to those skilled in the art ofpenicillin and cephalosporin chemistry. Methods for preparing certain ofthe more preferred easily cleavable esters, i.e. the pivaloyloxymethyl,acetoxymethyl, methoxymethyl, acetonyl and phenacyl esters, aredisclosed in U.S. Pat. No. 3,284,451 and in U.K. Patent 1,229,453.Preparation of phthalidyl esters of penicillins and cephalosporins isdescribed in South African Patent Applications 72/3799 and 72/3800. Thefree acid form of intermediate III may also be converted to a silylester, e.g. trimethylsilyl ester, as by the methods described in theliterature, e.g. U.S. Pat. No. 3,249,622. The silyl estercarboxylprotecting group may be easily removed following the acylationreaction by hydrolysis or alcoholysis.

Prior to the acylation reaction, any reactive substituents on theacylating acid or derivative thereof, e.g. hydroxy, carboxyl ormercapto, may be protected by use of suitable protecting or blockinggroups which are well-known to those skilled in the art of β-lactamchemistry, e.g. as by acylation or silylation. When the acylating agentcontains an amino functional group in the acyl moiety, the amino groupis protected by a conventional amino-blocking group which may be readilyremoved at the conclusion of the reaction. Examples of suitableamino-protecting or blocking groups include t-butoxycarbonyl,carbobenzyloxy, 2-hydroxy-1-naphthcarbonyl, trichloroethoxycarbonyl,2-ethoxycarbonyl-1-methylvinyl and 2-methoxycarbonyl-1-methylvinyl. Aparticularly valuable amino-blocking group is a proton, as in theacylating agent of the formula ##STR98## Preferred amino-protectinggroups are t-butoxycarbonyl, carbobenzyloxy, the proton and a β-diketoneor β-ketoester as in U.K. Patent 1,123,333 or U.S. Pat. Nos. 3,325,479and 3,316,247, e.g. methyl acetoacetate, or a β-ketoamide as in Japan71/24714. When the t-butoxycarbonyl, carbobenzyloxy, β-ketoester,β-diketone or β-ketoamide protecting groups are employed, it ispreferred to convert the acylating acid containing the blocked aminogroup to a mixed anhydride, e.g. with ethyl or isobutyl chloroformate,before reaction with compound III or a salt thereof. After the acylationcoupling reaction, the amino-protecting group and any other functionalprotecting groups used may be removed by methods known per se to formthe desired product of formula II. With respect to amino-protectinggroups, the t-butoxycarbonyl group may be removed by use of formic acid,the carbobenzyloxy group by catalytic hydrogenation, the2-hydroxy-1-naphthcarbonyl group by acid hydrolysis, thetrichloroethoxycarbonyl group by treatment with zinc dust in glacialacetic acid, the proton by neutralization, etc.

Acylation of a free amino group of a cephalosporin or penicillin nucleusis a well-known reaction, and any of the functional equivalents of thecarboxylic acid RCOOH commonly used in penicillin or cephalosporinchemistry as acylating agents for primary amino groups may be employedin acylating intermediate III. Examples of suitable acylatingderivatives of the free acid include the corresponding acid anhydrides,mixed anhydrides (e.g., alkoxyformic anhydrides), acid halides, acidazides, active esters and active thioesters. The free acid may becoupled with compound III after first reacting said free acid withN,N'-dimethylchloroformininium chloride [cf. Great Britain, 1,008,170and Novak and Weichet, Experientia XXI, 6, 360(1965)] or by the use ofenzymes or of an N,N'-carbonyldiimidazole or an N,N'-carbonylditriazole[cf. South African Specification 63/2684] or a carbodiimide reagent[especially N,N'-dicyclohexylcarbodiimide, N,N'-diisopropylcarbodiimideor N-cyclohexyl-N'-(2-morpholinoethyl)carbodiimide: cf. Sheehan andHess, J.A.C.S., 77, 1967 (1955)], or of alkylylamine reagent [cf. R.Buijle and H. G. Viehe, Angew. Chem. International Edition, 3, 582,(1964)] or of an isoxasolium salt reagent [cf. R. B. Woodward, R. A.Olofson and H. Mayer, J. Amer. Chem. Soc., 83, 1010 (1961)], or of aketenimine reagent [cf. C. L. Stevens and M. E. Munk, J. Amer. Chem.Soc., 80, 4065 (1958)] or of hexachlorocyclotriphosphatriazine orhexabromocyclotriphosphatriazine (U.S. 3,651,050) or ofdiphenylphosphoryl azide [DPPA; J. Amer. Chem. Soc., 94, 6203-6205(1972)] or of diethylphosphoryl cyanide [DEPC; Tetrahedron Letters No.18, pp. 1595-1598)] or of diphenyl phosphite [Tetrahedron Letters No.49, pp. 5047-5050 (1972)]. Other examples of suitable amide couplingreagents which have been described in the literature include (CH₃)₂ SCH₂CCHBr/DMSO (J. Chem. Soc. (C) 1904 (1969), HCCOCH₃ (Rec. Trav. Chim. 74,769 (1955), (CH₃)₂ C(OCH₃)₂ (Chim. Ther. 2, 195 (1967), SiCl₄ (J. Org.Chem. 34, 2766 (1969), TiCl.sub. 4 (Can. J. Chem. 48, 983 (1970),(PNCl₂)₃ (J. Org. Chem. 33, 2979 (1968), SO₃.sup.. DMF (J. Org. Chem.24, 368 (1959), ion exchange resins (Helv. 44, 1546 (1961) and J.C.S. C,874 (1969) and ##STR99## (J. Chem. Soc. 4650 (1964). An equivalent ofthe acid chloride is the corresponding azolide, i.e., an amide of thecorresponding acid whose amide nitrogen is a member of a quasi-aromaticfive membered ring containing at least two nitrogen atoms, i.e.,imidazole, pyrazole, the triazoles, benzimidazole, benzotriazole andtheir substituted derivatives. As an example of the general method forthe preparation of an azolide, N,N'-carbonyldiimidazole is reacted witha carboxylic acid in equimolar proportions at room temperature intetrahydrofuran, chloroform, dimethylformamide or a similar inertsolvent to form the carboxylic acid imidazolide in practicallyquantitative yield with liberation of carbon dioxide and one mole ofimidazole. Dicarboxylic acids yield diimidazolide. The by-product,imidazole, precipitates and may be separated and the imidazolideisolated, but this is not essential. A preferred acylating agent forpreparing 7-acylamido compounds containing an α-amino substituent, e.g.α-aminobenzyl, α-amino-α-thienylmethyl, etc. is the N-carboxy anhydride(Leuch's anhydride). In this structure the group which activates thecarboxyl group also serves to protect the amino group. Another preferredacylating agent for introducing a side chain containing an α-aminofunctional group is the acid chloride hydrochloride, of the formula##STR100## which also serves a dual function of carboxyl activation andamino protection. Mention was made above of the use of enzymes to couplethe free acid with compound III. Included in the scope of such processesare the use of an ester, e.g. the methyl ester, of that free acid withenzymes provided by various microorganisms, e.g. those described by T.Takahashi et al., J.A.C.S., 94(11), 4035-4037 (1972) and by T. Nara etal., J. Antibiotics (Japan) 24(5), 321-323 (1971) and in U.S. Pat. No.3,682,777. A particularly preferred coupling agent for coupling theacylating acid with compound III (or a salt or ester thereof) isN-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ) as described inJ.A.C.S. 90, 823-824 and 1652-1653 (1968) and U.S. Pat. No. 3,455,929.

The particular process conditions, e.g. temperature, solvent, reactiontime, etc. selected for the coupling reaction are determined by thenature of the reactants and acylation method used and are known to thoseskilled in the art.

The acylating agents which may be used to form the N-acyl compounds offormula II are known in the literature along with methods for theirsynthesis or are disclosed in the examples which follow. In those caseswhere the acylating agent contains one or more asymmetric carbon atomsand thus exists in optically active forms, the compounds obtained usingsuch an acylating agent are ordinarily obtained in racemic form. Whenthe separate optical isomers are desired, the acylating agent can beresolved in a conventional manner such as by reacting the free acid withcinchonine, strychnine, brucine or the like, fractionally crystallizingto separate the diastereoisomeric salts and separately acidifying thesolid phase and the liquid phase to liberate the optical isomers.

The 7-acylamido compounds of the present invention may be isolated inany of the ways customarily employed for the isolation of correspondingcephalosporin compounds. Formation of a desired pharamaceuticallyacceptable carboxylic acid or acid addition salt is carried out by knownmethods, e.g. reaction of the acid of compound II (or ester in the caseof acid addition salts) with an appropriate base or acid.

A compound of the formula II in the form of the free acid or a saltthereof may be converted to a pharmaceutically acceptable salt thereofor to a physiologically hydrolyzed ester or pharmaceutically acceptablesalt thereof. Similarly, the product of formula II in the form of aneasily cleavable ester or salt thereof may be converted to the free acidproduct or a pharmaceutically acceptable salt thereof by removal of theesterifying group to form the free acid, e.g. by acidic or alkalinehydrolysis, by enzymatic hydrolysis (as with human or animal serum), byhydrogenolysis or by treatment with chemical reagents known to removeparticular blocking groups, e.g. sodium thiophenoxide as in U.S. Pat.No. 3,284,451, and subsequent treatment of the free acid with an acid orbase to form a pharmaceutically acceptable salt.

The easily cleavable esters of the compounds of formula II are useful asintermediates in the production of the free acid product. Thepivaloyloxymethyl, acetoxymethyl, phthalidyl, indanyl and methoxymethylesters are also useful as active antibacterial agents since an oraladministration they are rapidly hydrolyzed to the active metabolite.These esters are of particular interest because they provide an oraladministration different rates and amounts of absorption and givediffering concentrations of the active antibacterial agent in blood andtissues.

The 7-amino intermediates of general formula III may be prepared byselectively reducing a 7-azido intermediate of the formula ##STR101##wherein Z is optionally substituted C₁ -C₆ alkyl, aryl, aralkyl orheterocyclic and R" is an easily cleavable ester carboxyl-protectinggroup. The carboxylprotected compound may, if desired, be cleaved toproduce the free-acid intermediate III which can be converted to a saltby methods known per se.

Preferred reducing agents for use in preparing the intermediates offormula III include chemical reducing agents such as zinc and ammoniumchloride, aluminum amalgam and hydrogen sulfide in the presence of abase, e.g. triethylamine or ammonia. Catalytic hydrogenation may also beemployed with such catalysts as noble metals, preferably platinum orpalladium incluing derivatives thereof such as oxides, hydroxides andhalides, or Raney nickel, said catalysts being optionally supported on aconventional carrier such as carbon or diatomaceous earth. Catalytichydrogenation is performed with a non-reducible inert solvent, e.g.methanol, ethanol or ethyl acetate, and preferably at atmospheric orslightly elevated pressure at room temperature.

Compound III in the carboxyl-protected form or a salt thereof may beused directly as a starting material in the N-acylation processdiscussed above. Alternatively, the protected intermediate may bede-blocked to form the free carboxylic acid which may then be optionallyconverted to a salt or to another carboxyl-protected form, e.g. aphysiologically hydrolyzed ester or salt thereof. By proper selection ofreduction conditions and protecting groups, azido intermediate IV' maybe converted either simultaneously or in stepwise fashion to the 7-aminofree acid III. Thus, if mild hydrogenation conditions are used, e.g.catalytic hydrogenation with 10% Pd-on-charcoal or a mild chemicalreducing agent such as H₂ S in the presence of a base such astriethylamine or ammonia, the azido group may be reduced withoutconcomitant removal of esters resistant to such conditions, e.g. benzylor p-nitrobenzyl. If stronger reducing conditions are used such as 30%Pd-on-diatomaceous earth, both the azido group and most reducible esterswill be simultaneously reduced.

A preferred embodiment of the present invention is the processcomprising the consecutive steps of

1. selectively reducing a 7-azido intermediate of the formula IV' toproduce a carboxyl-protected 7-amino intermediate of formula III and, ifdesired, removing the carboxyl-protecting group to produce thecorresponding free acid intermediate of formula III or optionally a saltthereof; and

2. N-acylating intermediate III or a salt thereof with an acylating acidof the formula R-COOH where R is an acyl group, or with its functionalequivalent as an acylating agent for a primary amine and, if desired,(a) when R" is a carboxylprotecting group, converting the 7-acylatedester to the free acid compound or a physiologically hydrolyzed ester ora pharmaceutically acceptable salt of said acid or ester, or (b) when R"is hydrogen, converting the 7-acylated carboxylic acid to aphysiologically hydrolyzed ester or a pharmaceutically acceptable saltof said acid or ester and, if desired, resolving a resulting isomermixture into its component isomers.

The 7β-azido intermediates IV' may be prepared by two alternativemethods. In one procedure a dihalide intermediate of the formula##STR102## wherein Y' represents a displaceable leaving group,preferally a group such as halo or sulfonyloxy, e.g., alkyl- orsubstituted alkylsulfonyloxy or aryl- or substituted arylsulfonyloxy,and most preferally a group selected from halo, --OSO₂ -- (lower)-alkylincluding especially --OSO₂ CH₃, --OSO₂ CF₃ and --OSO₂ C₆ H₄ CH₃ (para),X and X' which may be the same or different each represent a halogenatom, preferally bromine or iodine and most preferally iodine and R' isan easily cleavable ester carboxylprotecting group is reacted in aninert organic solvent in the presence of an acid acceptor with anucleophile of the formula

Z-SH

wherein Z is optionally substituted C₁ -C₆ alkyl, aryl, aralkyl orheterocyclic or a salt thereof, to form a thiolated intermediate of theformula ##STR103## Intermediate VI is then converted to the desired7β-azido compound by cyclization with base in an inert organic solvent.

The dihalide starting material V may be used in either of its isomericforms ##STR104## or as a mixture of isomers. Formula V above is intendedto represent either of the individual isomers or the mixture. Anydihalide including a mixed dihalide, e.g. X = Cl, X' = Br, may be usedbut the most preferred compound is the diiodide. Compound V is reactedin an inert organic solvent, e.g., methylene chloride, with the desiredthiol or a salt thereof. Since an acid HX is given off during thereaction, an acid acceptor, preferally an organic base such as pyridineor a trialkylamine, is used. Compound V, the thiol and the acid acceptorare preferally employed in approximately equimolar amounts. Thetemperature for the displacement step is not critical, but best resultsare obtained at room temperature or below, most preferally at atemperature of about 0° C.

Cyclization of intermediate VI is carried out in an inert organicsolvent, preferably a polar organic solvent such as dimethylsulfoxide ordimethylformamide, with a suitable base. The base used in thecyclization step may be selected from a wide variety of bases includingespecially those of the following categories:

a. anions derived from carboxylic acids having a pK_(a) of between about3.5 and 5.5;

b. tertiary organic amines such as a trialkylamine (e.g.,triethylamine), pyridine, n-methylpiperidine, n-methylmorpholine, etc.;

c. alkali metal hydrides, e.g., sodium or potassium hydride; and

d. organolithium compounds including especially lithium alkyls, e.g.,methyl, lithium or butyl lithium.

Most preferred cyclization bases are acetate and formate anions, e.g.,from alkali metal, ammonium or substituted ammonium formates oracetates. The most preferred base is acetate anion. The base ispreferally used in a molar excess relative to compound VI. While thepreferred temperatures for this step are room temperature or below, thetemperature is not critical. The leaving group Y' in formula VI shouldbe one which is efficiently displaced under the conditions of the basecyclization reaction and is preferally halo (chloro, bromo, fluoro, oriodo) or sulfonyloxy. The most preferred leaving group is the mesylategroup.

Preparation of intermediate V is described in U.S. Application Ser. No.567,323 filed Apr. 11, 1975 and also in the text below under the heading"Preparation of Starting Materials". Briefly summarized, the reactionscheme is as shown in Flow Sheet I below: ##STR105##

An alternative method for preparing the 7β-azido intermediates offormula IV' involves reacting an intermediate of the formula ##STR106##wherein Y" represents a displaceable leaving group and R' is an easilycleavable ester carboxyl-protecting group in an inert organic solvent inthe presence of an acid acceptor with a thiol of the formula

HS-Z

wherein Z represents an optionally substituted C₁ -C₆ alkyl, aryl,aralkyl or heterocyclic group, or a salt thereof.

Leaving group Y" in formula VII may be any nucleophilic group which isdisplaceable by the thiol. Examples of suitable leaving groups includehalo (chloro, bromo, fluoro or iodo) and sulfonyloxy, i.e., alkyl- orsubstituted alkylsulfonyloxy or arylor substituted arylsulfonyloxy.Preferred leaving groups are halo, --OSO₂ -(lower)alkyl includingespecially --OSO₂ CH₃, --OSO₂ CF₃ and --OSO₂ C₆ H₄ CH₃ (para). A mostpreferred leaving group is the mesylate group.

The nucleophilic displacement reaction is conducted in an inert organicsolvent, e.g., methylene chloride, in the presence of an acid acceptor,preferally an organic base such as pyridine or a trialkylamine. Theheterocyclic thiol, acid acceptor and intermediate VII are preferallyused in approximately equimolar quantities. The temperature is notcritical, but best results are obtained at room temperature or below,most preferally at a temperature of about 0° C.

The preferred starting materials of formula VII i.e., Y" = halo orsulfonyloxy, used in the above process are disclosed and claimed in U.S.application Ser. No. 567,323 filed Apr. 11, 1975; the entire disclosureof said application being herein incorporated by reference. A preferredmethod of preparation of these starting materials involves the steps of

1. cyclizing in an inert organic solvent, e.g., dimethylformamide, adihalide intermediate of formula V with a base R₂ COO⁻ in which R₂ inhydrogen or (lower) alkyl to produce an intermediate of the formula##STR107## in which R₂ is hydrogen or (lower) alkyl and; 2. subjectingsaid intermediate to acid hydrolysis to produce an intermediate of theformula ##STR108## either a. esterifying the 3-hydroxymethylintermediate with a sulfonic acid derivative, e.g., methanesulfonylchloride, p-toluenesulfonyl chloride or triflic anhydride, in thepresence of an organic base and in an inert organic solvent, e.g.,methylene chloride, to produce the desired 3-sulfonyloxy derivative offormula VII; or

b. halogenating the 3-hydroxymethyl intermediate with a phosphorushalide, e.g., phosphorus trichloride, or phosphorus tribromide, toproduce the desired 3-halomethyl derivative of formula VII.

The present invention also provides an alternative method for preparingthe novel 7-acylamido compounds of formula II; which process comprisesreacting in an inert organic solvent in the presence of an acid acceptora 7-acylated compound of the formula ##STR109## wherein R is an acylgroup, Y" is a displaceable leaving group and R" is hydrogen or aneasily cleavable ester carboxyl-protecting group, or a salt thereof,with a thiol of the formula

HS-Z

wherein Z is optionally substituted C₁ -C₆ alkyl, aryl, aralkyl orheterocyclic, or a salt thereof and, if desired, (a) when R" is acarboxyl-protecting group, converting the 7-acylated ester to the freeacid compound or a physiologically hydrolyzed ester or apharmaceutically acceptable salt of said acid or ester, or (b) when R"is hydrogen, converting the 7-acylated carboxylic acid to aphysiologically hydrolyzed ester or a pharmaceutically acceptable saltof said acid or ester; and, if desired, revolving a resulting isomermixture into its component isomers.

The nucleophilic displacement of the Y" group in compound VIII may becarried out using the same general reaction conditions, i.e., solvents,temperature range, acid acceptors, as described above in connection withthiolation of compound V.

The 7-acylated starting materials for the above process may be preparedby the procedures disclosed in U.S. application Ser. No. 567,323 filedApr. 11, 1975. While Y" may be any nucleophilic leaving group which isdisplaceable by the desired thiol, the preferred starting materials arecompounds wherein Y" is halo (chloro, bromo, iodo or fluoro) orsulfonyloxy, i,e., alkyl- or substituted alkylsulfonyloxy or aryl- orsubstituted arylsulfonyloxy. Examples of preferred leaving groupsinclude halo, --OSO₂ -(lower)alkyl, --OSO₂ CF₃ and --OSO₂ C₆ H₄ Ch₃(para). A most preferred leaving group is the mesylate group.

Starting materials of formula VIII may be prepared from 7β-azidocompounds of formula VII by the consecutive steps of

1. Selectively reducing intermediate VII as by catalytic hydrogenationwith a noble metal catalyst, e.g., 10% Pd-on-charcoal, or by use ofchemical reducing agents such as zinc and ammonium chloride, aluminumamalgam or hydrogen sulfide in the presence of a base such as ammonia ortriethylamine to produce a 7-amino intermediate of the formula##STR110## 2. N-acylating said 7-amino intermediate or a salt thereofwith an acylating acid of the formula

R-COOH

wherein R is an acyl group, or with its functional equivalent as anacylating agent for a primary amine, and, if desired, converting thecarboxyl-protected 7-acylamino compound to the corresponding free acidor to a salt thereof as by the methods disclosed above in connectionwith the de-blocking of compound II.

The pharmaceutically active compounds of the present invention arepotent antibacterial agents useful in the treatment of infectiousdiseases in poultry and animals, including man, caused by manyGram-positive and Gram-negative bacteria. The active compounds are alsoof value as nutritional supplements in animal feeds and as agents forthe treatment of mastitis in cattle.

The novel medicaments provided by the present invention may beformulated as pharmaceutical compositions comprising, in addition to theactive ingredient, a pharmaceutically acceptable carrier or diluent. Thecompounds may be administered both orally and parenterally. Thepharmaceutical preparations may be in solid form such as capsules,tablets or dragees, or in liquid form such as solutions, suspensions oremulsions. In the treatment of bacterial infections in man, the activecompounds of this invention may be administered parenterally or orallyin an amount of from about 5 to 200 mg/Kg./day and preferably about 5 to20 mg./Kg./day in divided dosage, e.g. three or four times a day. Theyare administered in dosage units containing, for example, 125, 250 or500 mg. of active ingredient with suitable physiologically acceptablecarriers or excipients.

Illustrative examples of the preparation of starting materials andcompounds of the present invention follow. These examples are given inillustration of, but not in limitation of, the present invention. Alltemperatures are in degrees Centigrade. DMF representsdimethylformamide, THF stands for tetrahydrofuran and EEDQ is the amidebond forming reagent having the structure ##STR111##

The β-lactam compounds prepared in the examples which follow all havethe hydrogen atoms at carbons 6 and 7 cis with respect to each otherand, unless indicated, the products are racemic mixtures in the sensethat they are composed of equal parts of the two isomers having thefollowing structures: ##STR112##

PREPARATION OF STARTING MATERIALS Preparation 1 ##STR113##

The procedure was essentially the same as that used to make thecorresponding ethyl ester by H. Adkins and J. Reeve, JACS 60, 1328(1938).

In a three necked one liter flask fitted with a thermometer, a droppingfunnel and a magnetic stirrer were placed 173 g. (0.9 mole) of benzylacetoacetate [The benzyl acetoacetate was prepared as described by Bakeret al., J, Org. Chem 17, 91 (1952)] and 130 ml. of glacial acetic acid.The contents were cooled in an ice bath and a solution of 69 g. (1 mole)of sodium nitrite in 130 ml. of water was added over a period of half anhour; the temperature was kept at 0 to 10° C. After the reaction mixturewas stirred for one hour at room temperature, 400 ml. of water was addedand the stirring was continued for an additional two hours. The reactionmixture was extracted three times with 200 ml. portions of diethylether. The diethyl ether extracts were combined, washed once with water,three times with saturated sodium bicarbonate solution and once withbrine. After drying over anhydrous sodium sulfate, the diethyl ethersolution was evaporated leaving [1.1] as a clear oil which solidifiedupon trituration with petroleum ether (30°-60° ) to give 186.5 g.(93.2%) of white solid. Its NMR spectrum was consistent with theassigned structure. Generally the product was used as such in subsequentreaction but it can be recrystallized from toluene, m.p. 81°-82° C.

Benzyl Oximino-Acetoacetate Ethylene Ketal ##STR114##

In a two liter flask fitted with a Dean Stark water separator and acondenser were placed 186.5 g. (0.85 mole) of benzyloximino-acetoacetate [1.1], 62 g. (1 mole) of ethylene glycol, 800 ml.of benzene (reagent grade) and 2 g. (10.5 mmole) of p-toluenesulfonicacid monohydrate. The reaction mixture was boiled at reflux until 15 ml.of water was removed (3 hours). The benzene solution was washed oncewith saturated sodium bicarbonate solution and once with brine. Afterdrying over anhydrous sodium sulfate, the benzene solution wasevaporated, leaving 212 g. (94% yield) of benzyl oximinoacetoacetateethylene ketal [2.1] as a light yellow oil. Its NMR spectrum wasconsistent with the assigned structure. Generally, the product was usedas such in subsequent reactions but one of the isomers can becrystallized² from toluene-petroleum either (b.p. 30°-60° C.); m.p. 52°C.

Anal. Calc'd. for C₁₃ H₁₅ NO₅ : C, 58.86; H, 5.70; N, 5.28. Found: C,58.97; H, 5.68; N, 5.12.

1. a mixture of the syn and anti isomers.

2. Only 35% of the oil could be crystallized.

Benzyl Amino-Acetoacetate Ethylene Ketal ##STR115##

Freshly prepared aluminum amalgam¹ (from 27 g. of aluminum foil) in athree-necked one liter flask was covered with 500 ml. of diethyl ether.The flask was fitted with a mechanical stirrer, a condenser, and adropping funnel. A solution of benzyl oximino-acetoacetate ethyleneketal [2.1] (132.5 g.; 0.5 mole) in 300 ml. of wet diethylether² wasadded dropwise at such a rate as to maintain boiling at reflux. Afterstirring for four hours, the reaction mixture was filtered through aBuchner funnel. The filtrate was evaporated leaving 110 g. of yellowishoil. The oil was dissolved in 800 ml. of dry diethylether and dryhydrogen chloride gas was bubbled into the solution until no furtherprecipitation occurred. The white precipitate was filtered off andwashed once with diethylether and then dried in vacuo. This provided 108g. of benzyl aminoacetoacetate ethylene ketal hydrochloride³ [3.1]; m.p.157-158° C.

Anal. Calc'd. for C₁₃ H₁₇ NO₄.HCl: C, 54.26; H, 6.31; N, 4.87. Found: C,53.96; H, 6.19; N, 4.60.

To obtain the free base, the hydrochloride salt was suspended in 500 ml.of diethylether and concentrated ammonium hydroxide was added withshaking until the solid went into solution. The diethylether layer wasseparated and washed twice with brine. After drying over anhydroussodium sulfate, the solvent was evaporated leaving 90 g. (71% yield) ofcolorless oil.

1. The aluminum amalgam was prepared essentially as described in A.I.Vogel ("Practical Organic Chemistry", 3rd, Edn., Longemans Green & Co.,London, (1957), p. 198) except for the following modification:

a. 5% NaOH was used.

b. The second washing with ethanol was omitted.

c. Dry diethylether was used for washing and most of the water must bedrained.

2. The diethylether was saturated with water by shaking with water in aseparatory funnel.

3. The product can be stored as the hydrochloride salt.

Schiff Base Formation And β-Lactam Formation ##STR116##

In a one liter flask fitted with a Dean Stark water separator and acondenser were placed 70.3 g. (0.28 mole) benzyl aminoacetoacetateethylene ketal [3.1], 37 g. (0.28 mole) cinnamaldehyde, and 750 ml. ofmethylene chloride (reagent grade). The mixture was boiled at reflux for30 minutes and then 400 ml. of methylene chloride were distilled off.The concentrated solution was then dried over anhydrous sodium sulfateand then evaporated to dryness in vacuo¹. The residual oil was checkedby NMR to ensure that Schiff base formation was complete beforecontinuing on to the next step.

The freshly prepared Schiff base [4.1] was diluted with 600 ml. ofmethylene chloride² and cooled to 0° C. (ice-salt bath). Triethylamine(31.1 g.; 0.308 mole) was added and then a solution of 36.2 g. (0.308mole) of azidoacetyl chloride³ in 362 ml. of methylene chloride² wasadded dropwise at 0° C. over a period of 1 hour. The reaction mixturewas stirred for an additional hour at room temperature.sup. 4 and thenevaporated on a rotary evaporated at reduced pressure while being heatedon a 35° C. water bath⁵. The residue was diluted with 500 ml. ofdiethylether and filtered. The filtrate was washed twice with brine anddried over anhydrous sodium sulfate. Evaporation of this solutionyielded 117.5 g. (94% yield) of styryl β-lactam [5.1]. Its NMR and IRspectra are consistent with the assigned structure and indicate thepresence of a mixture of isomers, diasteriomeric at the carbon α to thecarbonyl of the benzyl ester. 1. This evaporation must be done to ensurecomplete Schiff base formation.

2. All the methylene chloride used in the cycloaddition reaction wasreagent grade which was first dried over molecular sieve (Type 4A) andthen over anhydrous calcium chloride. It was stored thereafter overmolecular sieve (Type 4A).

3. j. h. boyer and J. Horner, J. Amer. Chem. Soc., (1955), 77, 951.

4. The reaction mixture can be kept overnite at 0° if necessary.

5. This operation is necessary to ensure complete β-lactam formation.##STR117##

Styryl β-lactam [5.1](117.5 g.; 0.262 mole) was dissolved in one literof methylene chloride (reagent grade), cooled to -50° to -60° C. in adry ice-acetone bath, and ozonized until a faint blue-green colorappeared. The solution was then flushed with nitrogen until the colorfaded. Methylsulfide (100 ml.) was added to the -50° C. solution, whichwas then allowed to slowly reach 25° as the cooling bath graduallymelted. It was kept overnite at room temperature under nitrogen and thenit was washed twice with 1% sodium bicarbonate solution, twice withbrine, dried over anhydrous sodium sulfate, and evaporated to dryness.The resulting oil triturated four times with 100 ml. portions ofpetroleum ether (b.p. 30-60° C.) to remove benzaldehyde. The oil wasthen triturated carefully with diethylether whereupon it solidified. Thesolid was filtered off and dried to provide 75 g. (71.5%) of aldehyde[6.1] as a mixture of isomers diasteriomeric at the carbon α to thecarbonyl of the benzyl ester. Recrystallization of [6.1] from ether gavewhite crystals, m.p. 101°-102° C. (corrected).

Anal. Calc'd. for C₁₇ H₁₈ N₄ O₆ : C, 54.54; H, 4.84; N, 14.96. Found: C,54.75; H, 4.87; N, 14.89. ##STR118##

The aldehyde [6.1] (116.3 g.; 0.31 mole) was dissolved in 600 ml. of THF(reagent grade) and the solution was then cooled to -10° C.(ice-methanol bath). Sodium borohydride (5.88 g.; 0.155 mole) was addedand the reaction mixture was stirred 1 hour. 10% aqueous hydrochloricacid was added until the mixture was slightly acidic, then 600 ml. brinewas added. The THF layer was separated and the aqueous phase wasextracted twice with 250 ml. portions of diethylether. The combinedorganic phases were washed twice with 400 ml. portions of brine, driedover anhydrous sodium sulfate, and evaporated in vacuo to yield 117.3 g.of crude alcohol [7.1] as an orange oil. This oil was used as such inthe next reaction.

A solution of methanesulfonyl chloride (37.8 g.; 0.34 mole) in 100 ml.of methylene chloride.sup. 1 was added dropwise at 0° C. (ice-waterbath) to a stirring solution of alcohol [7.1] (105.6 g.; 0.28 mole,triethylamine (56.6 g.; 0.34 mole) and one liter of methylene chloride¹.Afterwards, the reaction was stirred for 30 hours at 25° C. It was thenwashed twice with brine (500 ml. portions), dried over anhydrous sodiumsulfate, and evaporated in vacuo. The resulting oil was dissolved inmethylene chloride, treated with norite, and then filtered overapproximately 200 g. of activity I silica gel. The silica gel was thenwashed with approximately 2 liters of methylene chloride. The filtratewas evaporated to dryness and the resulting oil (116 g.) was coveredwith diethylether. It crystallized on standing to give 87.2 g. (80% from[6.1] of mesylate [8.1] as off-white solid, m.p. 97°-99° C. (corrected).

1. The methylene chloride used was reagent grade which had been furtherpurified by passing over a column of calcium chloride and then storingover molecular sieve (Type 4A). ##STR119##

A mixture of mesylate [8.1] (3.19 g.; 6.43 mmole) and 30 ml. of 95%trifluoroacetic acid was stirred at 25° for 2 hours. The mixture wasdiluted with 300 ml. of brine and extracted three times with methylenechloride (100 ml. portions). The combined extracts were washed threetimes with water (50 ml. portions, until neutral), dried (anhydroussodium sulfate) and evaporated to dryness in vacuo leaving 3.17 g. of abrown oil. NMR spectra of this oil indicate the presence of >90% enol[9.1].

Crude enol [9.1] (48.0 g.; 0.117 mole) and triflic anhydride (33.0 g.;0.117 mole) were dissolved in 500 ml. of methylene chloride and thesolution was then cooled to 0° C. (ice-water bath). A solution oftriethylamine (11.8 g.; 0.117 mole) in 80 ml. of methylene chloride² wasadded dropwise over a period of 40 minutes. When the addition wascomplete, the ice-water bath was removed and the mixture was stirred at25° for 45 minutes. The mixture was then poured into 300 ml. of icewater and washed with cold water until the pH of the washings wasapproximately 6. The extract was dried (anhydrous sodium sulfate) andevaporated in vacuo to give 54.0 g. of crude triflate [10.1] as a darkred oil. This oil was dissolved in 400 ml. of benzene (USP) and passedthrough a 1 1/2 inch pad of activity III silica gel. The pad was washedwith 1 1. of benzene. Evaporation of the benzene gave 38.3 g. of ayellow oil. This oil was carefully triturated with 50 ml. of absoluteethanol and then cooled at 0° C. for 2 hours. The resulting white solidwas filtered off and dried in vacuo to give 19.5 g. of triflate [10.1]as one isomer, m.p. 57°-59° C. (corrected).

Anal. Calc'd. for C₁₇ H₁₇ F₃ N₄ O₄ S₂ : C, 37.67; H, 3.14; N, 10.33; S,11.82. Found: C, 37.40; H, 3.12; N, 10.43; S, 11.73.

1. triflic anhydride was prepared as follows: 170 g. (100 ml.) CF₃ SO₃ H("Fluorochemic acid" 3M Company) and 135 g. P₂ O₅ were mixed carefully,shaken well, and stored 18 hours protected from moisture. The productwas distilled from the resulting solid mass using a flame; the fractionboiling 8°-90° C. was collected. Re-distillation of this fractionyielded 119.45 g. (74%) of triflic anhydride boiling 82°-84° C.

2. the methylene chloride used was reagent grade which had been furtherpurified by passing over a column of calcium chloride and then storedover molecular sieve (Type 4A). ##STR120##

Triethylamine (1 g.; 0.01 mole) was added to a stirred solution oftriflate [10.1] (5.42 g.; 0.01 mole) in 55 ml. of methylene chloride(A.R.) at room temperature. After stirring for five minutes (at whichpoint TLC shows complete formation of allene[11.1], a solution ofbromine (10 ml. of 1M solution in CCl₄ ; 0.01 mole) was added dropwise.After addition of the bromine, the mixture was concentrated, absorbedonto Activity I silica gel and dry column chromatographed on Activity Isilica gel by eluting with methylene chloride (USP). This yielded onefraction (uniformly one spot by TLC) weighing 2.5 g. (45%). Its IR, UV,and NMR spectra were consistent with the expected dibromide structure[13.1].

Anal. Calc'd. for C₁₆ H₁₂ Br₂ N₄ O₆ S: C, 34.80; H, 2.92; N, 10.15.Found: C, 35.25; H, 2.97; N, 10.02. ##STR121##

A solution of triethylamine (101 mg., 1.00 mmole) in 1.4 ml. ofmethylene chloride was added with stirring to a solution of triflatefrom Preparation 1A above [10.1] (542 mg., 1.00 mmole) in 5.4 ml. ofmethylene chloride at 0° C. After allowing the solution to warm to 24°over 15 minutes, a solution of iodine (254 mg., 1.00 mmole) in 7.5 ml.of methylene chloride was added with stirring over 30 minutes, thenwashed with water, dried, decolorized, filtered and the solventevaporated in vacuo to give the diiodide [23.1] (588 mg.; 91% yield) ingreater than 95% purity. The IR and NMR spectra were consistent for theproposed structures.

Anal. Calc'd. for C₁₆ H₁₆ N₄ O₆ I₂ S: C, 29.74; H, 2.50; N, 8.67; I,39.28; S, 4.96. Found: C, 29.76; H, 2.47; N, 8.61; I, 39.37; S, 5.18.

Other suitable intermediates of general formula V in the specificationand claims may be prepared by:

1. substituting another easily cleavable ester group for the benzylester of starting material 1.1; or

2. esterifying compound 7.1 with another sulfonic acid derivative orhalogenating compound 7.1; or

3. halogenating allene 11.1 with a halogenating agent other than bromineor iodine, e.g. BrCl.

Preparation 2

p-Nitrobenzyl 7β-azido-3-methylsulfonyloxymethyl-Δ³-0-2-isocephem-4-carboxylate ##STR122##

A solution of methanesulfonyl chloride (0.50 ml., 6.5 mmole) in 10 ml.of methylene chloride was added dropwise with stirring to a solution ofp-nitrobenzyl 7β-azido-3-hydroxymethyl-Δ³ -0-2-isocephem-4-carboxylate(2.41 g., 6.43 mmole), triethylamine (0.97 ml., 7.0 mmole) and 75 ml. ofmethylene chloride at -10°. After 1/2 hour at -10° and 1 hour at 24°,the solution was washed with 5% hydrochloric acid, 2% sodiumbicarbonate, and water (85 ml. each), then the solvent was evaporated invacuo to give the mesylate title product, 2.86 g. (98% yield), as ayellow foam. The NMR of the product was in agreement with the proposedstructure.

The p-nitrobenzyl 7β-azido-3-hydroxymethyl-Δ³ -0-2-isocephem-4-carboxylate starting material used above may be preparedas follows:

1. The diiodide intermediate of the formula ##STR123## was prepared fromp-nitrobenzyl acetoacetate according to the procedures of Preparations1A and B (Starting Materials) described above.

2. The diiodide intermediate (6.6 g., 9.6 mmole) was cyclized withpotassium formate (2.54 g., 30 mmole) in a solution of 100 ml. DMF and0.1 ml. water at 0°. After stirring for 5 hours with the cooling bathremoved, the mixture was poured into 100 ml. of cold water and extractedwith methylene chloride. After washing with water containing a littleNaCl, drying and evaporation in vacuo, p-nitrobenzyl7β-azido-3-formyloxymethyl-Δ³ -0-2-isocephem-4-carboxylate was recovered(5.3 g.) as a brown oil.

3. To a solution of 5.3 g. of the 3-formyloxymethyl intermediate in 53ml. of acetone was added 26 ml. of water and 3.2 ml. of 12M HCl. Themixture was stirred at 24° for 7 hours, then poured into 100 ml. waterand extracted with methylene chloride. The combined extracts were washedwith water containing a little sodium chloride, dried and evaporated invacuo to give 3.6 g. of a brown oil. The oil was absorbed from methylenechloride onto 18 g. of silica gel and placed on a 72 g. silica gelcolumn (grade 3, 5% ether). The column was eluted with 200 ml. of ether,then with ether/ethyl acetate 3:1. The major component (Rf 0.20) gave,on evaporation of the solvent in vacuo, a yellow solid which wasrecrystallized from acetone-ether to give the 3-hydroxymethyl startingmaterial of this example, 950 mg. (17.5% yield from the diiodide). m.p.147°-148°.

Anal. Calc'd. for C₁₅ H₁₃ N₅ O₇ : C, 48.00; H, 3.49; N, 18.66. Found: C,48.11; H, 3.61; N, 18.81.

By replacement of the methanesulfonyl chloride in the procedure abovewith other sulfonic acid derivatives and the p-nitrobenzyl ester withother easily cleavable esters, suitable intermediates may be preparedhaving the formula ##STR124## in which Y" is sulfonyloxy and R' is aneasily cleavable ester residue.

Preparation 3

Benzyl 7β-azido- 3-bromomethyl-Δ³ -0-2-isocephem-4-carboxylate##STR125##

To a solution of benzyl 7β-azido-3-hydroxymethyl-Δ³-0-2-isocephem-4-carboxylate in benzene is added about an equimolaramount of pyridine and a slight molar excess of phosphorus tribromide.There is produced the 3-bromomethyl title product.

The benzyl 7β-azido-3-hydroxymethyl-Δ³ -0-2-isocephem-4-carboxylate usedabove may be prepared by the method disclosed in Preparation 2 bysubstituting the benzyl ester of the diiodide intermediate for thep-nitrobenzyl ester used therein.

Other suitable 7β-azido-3-halomethyl esters may be prepared by the abovemethod by appropriate variation of the ester acetoacetate startingmaterial and the halogenating agent.

Preparation 4

Benzyl 7β-phenoxyacetamido-3methyl-sulfonyloxymethyl-Δ³-0-2-isocephem-4-carboxylate ##STR126##

Benzyl isocephem-7β-azido-3-methylsulfonyl-oxymethyl-Δ³-0-2-isocephem-4-carboxylate (Preparation 2) is dissolved in methylenechloride and cooled to 0° C. Two equivalents of triethylamine are addedand, while stirring and cooling, H₂ S gas is passed through the solutionuntil it is saturated. The solution is allowed to come to room andconcentrated to give benzyl 7β-amino-3-methylsulfonyl-oxymethyl-Δ³-0-2-isocephem-4-carboxylate.

The ester is reacted with equimolar amounts of phenoxyacetic acid andEEDQ in methylene chloride to give the title product.

Other N-acylated compounds of the formula ##STR127## may be prepared bysubstituting for the phenoxyacetic acid used above another desiredacylating agent, preferally one which will produce a starting materialin which R is one of the acyl groups mentioned as being preferred inconnection with the novel end-products of formula II.

Preparation 5

1-carboxymethyl-1,2,3,4-tetrazole-5-thiol and its di-sodium salt##STR128## A. Recrystallization of 1-methyl-5-mercaptotetrazoleProcedure:

1. One hundred and ten grams of 1-methyl-5-mercaptotetrazole is slurriedin 350 ml. of boiling chloroform. A near solution is obtained.

2. The hot solution (50°-60°) is rapidly filtered by vacuum through aheated Buchner funnel (11 cm. SS No. 604 paper containing one-fourth toone-third inch of packed filter aid ("Supercel"). The filter pad iswashed with 50 ml. of 50°-70° C. chloroform which is added to thefiltrate.

3. The filtrate is cooled to approximately 0°-6° C. and kept at 0°-6° C.for 2 hours. The crystals which have formed are collected by filtrationat 0°-6° C. and washed with 60 ml. of 0°-6° C. chloroform which is addedto the filtrate. The crystals (cut A) are air dried at 37°-45° C. for 18hours.

4. The filtrate is concentrated on the rotary vacuum evaporator (60° C.bath) to approximately one-half volume. This slurry is cooled to 0°-6°C. and kept at 0°-6° C. for 2 hours. The crystals are collected byfiltration at 0°-6° C., washed with 40 ml. of 0°-6° C. chloroform whichis added to the filtrate. The crystals (cut B) are air dried at 37°-45°C. for 18 hours. Crystal cuts A and B are composited to give anapproximate 65% weight yield.

5. The filtrate of cut B, Step 4 may be reworked twice as described inStep 4 to obtain an additional 15% recovery.

B. preparation of 1-carboxymethyl-1,2,3,4-tetrazole-5-thiol and itsdi-sodium salt

1. Five hundred ml. of substantially dry and pure tetrahydrofuran in a2-liter 3 neck flask with stirrer is cooled in a salt-acetone-ice bathto approximately -10° C. Dry nitrogen gas is blown on the liquidsurface.

2. Five hundred ml. of 15.06% (1.6 N) butyl lithium in hexane (FooteMineral Co.) is added over a ten minute period under dry nitrogen andstirring to the tetrahydrofuran. The near solution is cooled to -5° to-10° C.

3. Forty six and four tenths gram (46.4 g.) of1-methyl-5-mercaptotetrazole (recrystallized as above) is dissolved in200 ml. of substantially pure and dry tetrahydrofuran. The solution isfiltered if cloudy and then cooled to 5° to 10° C.

4. The cooled solution of step 3 is added over 10 minutes with stirringand under dry nitrogen to the butyl lithium solution. The temperatureshould be maintained at -5° C. to +10° C. maximum. Precipitates mayform.

5. The mixture is stirred under dry nitrogen and 0° C. to +10° C. forone half hour.

6. Anhydrous carbon dioxide gas is bubbled through at a rapid rate andwith rapid stirring for 15-30 minutes at approximately ambienttemperature (0° to 10° C.) to no higher than +20° C.

7. The white precipitate which forms is suitably collected by filtrationin an area of low humidity. The precipitate is washed with about 75 ml.of tetrahydrofuran.

8. The precipitate is dissolved in 250 ml. of water (pH 8.5-9.5). Asecond layer of tetrahydrofuran may be present. This may be removed inthe vacuum rotary evaporator (50° C. bath).

9. The aqueous solution is adjusted to pH 1.6-2.0 with concentratedhydrochloric acid.

10. The acid aqueous solution is extracted twice with 250 ml. portionsof ethyl acetate. Each 250 ml. ethyl acetate extract is back extractedwith 100 ml. portions of water. The water extracts are discarded. Theethyl acetate extracts (free of any water layer) are filtered andcomposited.

11. The combined ethyl acetate extracts are concentrated to dryness onthe vacuum rotary evaporator (60° C. bath).

12. The crystals in the flask are boiled with 300 ml. of chloroform forabout 2 minutes. The hot slurry (50°-60° C.) is vacuum filtered througha heated Buchner funnel (11 cm-SS-604 paper). The crystals are washedwith about 75 ml. of 50° C. chloroform. The crystals are air dried atroom temperature for about 3 hours and then made about 100-200 mesh.

13. The 100-200 mesh crystals are treated with boiling chloroformexactly as described in step 12 (the hot chloroform removes most of theunreacted 1-methyl-5-mercaptotetrazole). Yield: approximately 45 to 50grams of crystalline 1-carboxymethyl-1,2,3,4-tetrazole-5-thiol. Thesecrystals may contain 0.02 to 0.05 moles of 1-methyl-5-mercaptotetrazole.

14. The crystals of step 13 are slurried with 250 ml. of ethyl ether atroom temperature for 3-5 minutes. The mixture is filtered. Theinsolubles (0.5-5%) may be a contaminating symmetrical mercaptotetrazoleketone of the following tentative structure: ##STR129## CAUTION: Thiscompound EXPLODES at approximately 205°-210° C.

15. The ether filtrate of step 14 is evaporated to dryness on the vacuumrotary evaporator (50° C. bath). Approximately 42 to 48 grams ofcrystalline 1-carboxymethyl-1,2,3,4-tetrazole-5-thiol containingapproximately 0.01-0.05 mole of 1-methyl-5-mercaptotetrazole isrecovered.

16. The crystals are dissolved in 420 ml. of absolute ethanol(approximately 100 mg./ml.). The solution is warmed to 50°-60° C.

17. To the hot solution of step 16, 310 ml. of a 41% sodium2-ethylhexanoate (SEH) solution in isopropanol is added with very rapidstirring over a 10 minute period. A crystalline precipitate forms. Themixture is slurried at 50°-60° C. for 20 minutes.

18. The mixture is filtered hot (50°-60° C.) through a heated Buchnerfunnel (11 cm-SS-No. 604 paper). The crystals are washed with 75 ml. of50° C. ethanol.

19. The ethanol damp crystals of step 18 are slurried in 200-300 ml. ofethanol. The slurry is passed through a 200 mesh screen. The slurry isheated to 50°-60° C. for 5 minutes with rapid stirring (unreacteddi-sodium 1-methyl-5-mercaptotetrazole is very soluble in hot ethanol).

20. The crystals are collected at 50°-60° C. on a 11 cm-SS No. 604 paperin a heated Buchner funnel. The crystals are washed with 75-100 ml. ofethanol and vacuum dried at 50°-60° C. for 24-48 hours. Yield: 40-48grams of di-sodium 1-carboxymethyl-1,2,3,4-tetrazole-5-thiol (free of1-methyl-5-mercaptotetrazole as observed by NMR).

Preparation 6

1-Carboxyethyl-1,2,3,4-tetrazole-5-thiol ##STR130## A.2-Carboethoxyethylisocyanate

β-alanine ethyl ester hydrochloride (93.6 g.), triethylamine (123.5g)and methylene chloride (400 ml) were mixed together and cooled to -10°C. Carbon disulfide (46.5 g) dissolved in 150 ml. of chloroform wasadded to the above solution during a two-hour period while keeping thetemperature at about -10° C. After the addition was complete, thetemperature was allowed to warm to 10° C. for about 10 minutes. Thesolution was again cooled to -10° C. and 66.3 g of ethyl chloroformatein 60 ml of chloroform was added dropwise over a 40-minute period withstirring. The temperature was allowed to rise to room temperature for 30minutes and again cooled to 0° C. an additional 61.6 g of triethylaminewas added at 0° C. and then the solution was stirred at room temperaturefor 3 hours.

The mixture was treated with water and the organic phase collected,washed with 2 × 250 ml of 2N HCl, then 2 × 250 ml of NaHCO₃, then 2 ×250 ml of water. The organic phase was dried over NA₂ SO₄ and thesolvent removed in vacuo to produce 93.7 g of an oil found to be thedesired product. The IR and NMR spectra were consistent with thestructure.

B. 1-Carboxyethyltetrazol-5-thiol

Sodium azide (29.7 g) was dissolved in 400 ml of water and heated to 60°C in a nitrogen atmosphere. 2-Carboethoxyethylisocyanate (46.9 g)dissolved in 50 ml of Skellysolve B (essentially n-hexane) was added tothe heated sodium azide solution. The solution was stirred for about 150minutes at about 70°-72° C., then cooled to 30° C. in an ice bath. 50%sodium hydroxide solution was added until the pH was 12. The mixture washeated for forth minutes at 70° C. and cooled to 15° C. in an ice bath.The pH was adjusted to 2 using conc. HCl and then extracted with ethylacetate (4 × 150 ml). The ethyl acetate extracts were washed with water,then dried over sodium sulfate. The solvent was evaporated in vacuo andthe product was collected as crystals from methylene chloride to yield19.5 g of title product.

Substitution in the procedure for the preparation of1-carboxyethyltetrazol-5-thiol for the β-alanine ethyl ester usedtherein of an equimolar quantity of an appropriately substituted aminoacid ester of 3 to 4 carbon atoms produces the corresponding 1-carboxy(C₃ -C₄ alkyl)tetrazol-5-thiol, e.g., 1-carboxypropyltetrazol-5-thioland 1-carboxybutyltetrazol-5-thiol.

EXAMPLES EXAMPLE 1

p-Nitrobenzyl 7β-azido-3-(1-methyltetrazol-5-ylthiomethyl)-Δ³-0-2-isocephem-4-carboxylate ##STR131##

To a solution of p-nitrobenzyl 7β-azido-3-methylsulfonyloxymethyl-Δ³-0-2-isocephem-4-carboxylate (4.53 g., 10 mmole) and triethylamine (1.4ml., 10 mmole) in 90 ml. of methylene chloride was added 10 mmole of1-methyltetrazole thiol. The solution was stirred at 24° C. for 16hours, then washed with 5% HCl and water (100 ml. each), dried and thesolvent evaporated in vacuo to give the product as a yellow oil. Uponrecrystallization from ethyl acetate, the title product was obtained in78% yield; m.p. 150°-152° C. The NMR was consistent with the proposedstructure.

EXAMPLE 2

p-Nitrobenzyl 7β-azido-3-(2-methyl-1,3,4-thiadiazol-5-ylthiomethyl)-Δ³-0-2-isocephem-4-carboxylate ##STR132##

The procedure of Example 1 is repeated except that the 1-methyltetrazolethiol used therein is replaced by 10 mmole of 2-methylthiadiazole thiol.The title product was isolated as a yellow oil in 95% yield. The NMR andIR spectra of the product were in agreement with the proposed structure.

EXAMPLE 3

p-Nitrobenzyl-7β-azido-3-(2-methyl-1,3,4-oxadiazol-5-ylthiomethyl)-.DELTA.³-0-2-isocephem-4-carboxylate ##STR133##

A solution of p-nitrobenzyl 7β-azido-3-methylsulfonyloxymethyl-Δ³-0-2-isocephem-4-carboxylate (1.36 g., 3.0 mmoles) and triethylamine(0.38 ml., 3.0 mmoles) in 50 ml. of dichloromethane was treated with2-methyl-1,3,4-oxadiazole-5-thiol (0.35 g., 3.0 mmoles). The reactionmixture was stirred at room temperature for 48 hours and then washedwith 10% HCl, water and brine. The solution was dried over sodiumsulfate and evaporated in vacuo giving crude product. The crude productwas purified by dry-column chromatography on silica gel (60 g., ActivityIII) eluting with 15% ethyl acetate in ether. There was obtained 1.0 g.of pure title product in 70% yield. The NMR spectrum was in agreementwith the proposed structure.

EXAMPLE 4

If the general procedures of Examples 1-3 are repeated using equimolarweights of 1,2,3-triazole-5-thiol,1-carboxymethyl-1,2,3,4-tetrazole-5-thiol and1-carboxyethyl-1,2,3,4-terazole-5-thiol, respectively, in place of thethiols used therein, there are produced p-nitrobenzyl7β-azido-3-(1,2,3-triazol-5-ylthiomethyl)-Δ³-0-2-isocephem-4-carboxylate, p-nitrobenzyl7β-azido-3-(1-carboxymethyl-1,2,3,4-tetrazol-5-ylthiomethyl)-Δ.sup.3-0-2-isocephem-4-carboxylate and p-nitrobenzyl7β-azido-3-(1-carboxyethyl-1,2,3,4-tetrazol-5-ylthomethyl)-Δ.sup.3-0-2-isocephem-4-carboxylate, respectively.

EXAMPLE 5

Repeating the general procedures of Examples 1-3, p-nitrobenzyl7β-azido-3-methylsulfonyloxymethyl-Δ³ -0-2-isocephem-4-carboxylate isreacted with the thiols listed below to produce the following7β-azido-3-thiolated compounds.

    __________________________________________________________________________    Thiol                   Product                                               __________________________________________________________________________                     ##STR134##                                                   methyl mercaptan   methyl                                                     ethyl mercaptan    ethyl                                                      butyl mercaptan    butyl                                                      pentyl mercaptan   pentyl                                                     2-aminoethyl mercaptan                                                                           2-aminoethyl                                               1-chloroethyl mercaptan                                                                          1-chloroethyl                                              2-hydroxyethyl mercaptan                                                                         2-hydroxyethyl                                             2-bromoethyl mercaptan                                                                           2-bromoethyl                                               2-nitroethyl mercaptan                                                                           2-nitroethyl                                               5-nitropentyl mercaptan                                                                          5-nitropentyl                                              3-cyano-n-propyl mercaptan                                                                       3-cyano-n-propyl                                           4-(dimethylamino)-n-butyl                                                                        4-(dimethylamino)-n-butyl                                   mercaptan                                                                    2,3-dihydroxypropyl                                                                              2,3-dihydroxypropyl                                         mercaptan                                                                    3-chloro-2-methylbutyl                                                                           3-chloro-2-methylbutyl                                      mercaptan                                                                    benzenethiol       phenyl                                                     p-chlorobenzenethiol                                                                             p-chlorophenyl                                             2,4,5-trichlorobenzenethiol                                                                      2,4,5-trichlorophenyl                                      o-aminobenzenethiol                                                                              o-aminophenyl                                              p-bromobenzenethiol                                                                              p-bromophenyl                                              p-aminobenzenethiol                                                                              p-aminophenyl                                              2,5-dichlorobenzenethiol                                                                         2,5-dichlorophenyl                                         p-fluorobenzenethiol                                                                             p-fluorophenyl                                             m-methoxybenzenethiol                                                                            m-methoxyphenyl                                            p-methoxybenzenethiol                                                                            p-methoxyphenyl                                            p-nitrobenzenethiol                                                                              p-nitrophenyl                                              2,3,5,6-tetrachloroben-                                                                          2,3,5,6-tetrachlorophenyl                                   zenethiol                                                                    benzyl mercaptan   benzyl                                                     p-chlorobenzyl mercaptan                                                                         p-chlorobenzyl                                             phenethyl mercaptan                                                                              phenethyl                                                  p-nitrobenzyl mercaptan                                                                          p-nitrobenzyl                                              p-methoxybenzyl mercaptan                                                                        p-methoxybenzyl                                            1-naphthalenethiol 1-naphthyl                                                 2-naphthalenethiol 2-naphthyl                                                 4-chloro-1-naphthalenethiol                                                                      4-chloro-1-napthyl                                         4-nitro-1-naphthalenethiol                                                                       4-nitro-1-naphthyl                                         2-thienyl mercaptan                                                                              2-thienyl                                                  3-thienyl mercaptan                                                                              3-thienyl                                                  2-furyl mercaptan  2-furyl                                                    3-furyl mercaptan  3-furyl                                                    6-mercaptotetrazolo[4,5-6]                                                                       tetrazolo[4,5-6]pyridazin-6-yl                              pyridazine                                                                   3-hydroxy-6-mercaptopyrida-                                                                      3-hydroxypyridazin-6-yl                                     zine                                                                         6-bromo-3-pyridazinethiol                                                                        6-bromopyridazin-3-yl                                      3-pyridazinethiol  pyridazin-3-yl                                             3-methyl-1-phenyl-5-                                                                             3-methyl-1-phenylpyrazol-5-yl                               pyrazolethiol                                                                imidazole-2-thiol  imidazol-2-yl                                              1-methyl-5-nitro-2-imidazole-                                                                    1-methyl-5-nitroimidazol-2-yl                               thiol                                                                        thiazole-2-thiol   thiazol-2-yl                                               5-methylthiazole-2-thiol                                                                         5-methylthiazol-2-yl                                       oxazole-2-thiol    oxazol-2-yl                                                5-methyloxazole-2-thiol                                                                          5-methyloxazol-2-yl                                        2-pyridinethiol    2-pyridyl                                                  4-pyridinethiol    4-pyridyl                                                  3-amino-2-pyridinethiol                                                                          3-amino-2-pyridyl                                          5-nitro-2-pyridinethiol                                                                          5-nitro-2-pyridyl                                          3-methyl-1-phenyl-5-                                                                             3-methyl-1-phenyl-pyrazol-                                  pyrazolethiol      5-yl                                                      2-pyrazinethiol    pyrazin-2-yl                                               4-pyrimidinethiol  pyrimidin-4-yl                                             4-methyl-2-pyrimidinethiol                                                                       4-methylpyrimid-2-yl                                       3-methylisothiazole-5-thiol                                                                      3-methylisothiazol-5-yl                                    isothiazole-5-thiol                                                                              isothiazol-5-yl                                            1,2,3,4-thiatriazole-5-thiol                                                                     1,2,3,4-thiatriazol-5-yl                                   5-mercapto-3-methylthio-                                                                         3-methylthio-1,2,4-thiadiazol-                              1,2,4-thiadiazole  5-yl                                                      5-mercapto-3-methyl-1,2,4-                                                                       3-methyl-1,2,4-thiadiazol-                                  thiadiazole        5-yl                                                      2-mercapto-1,3,4-thiadiazole                                                                     1,3,4-thiadiazol-2-yl                                      5-mercapto-2-ethyl-1,3,4-                                                                        2-ethyl-1,3,4-thiadiazol-                                   thiadiazole        5-yl                                                      5-mercapto-2-n-butyl-1,3,4-                                                                      2-n-butyl-1,3,4-thiadiazol-                                 thiadiazole        5-yl                                                      5-mercapto-2-amino-1,3,4-                                                                        2-amino-1,3,4-thiadiazol-                                   thiadiazole        5-yl                                                      5-mercapto-2-methylamino-                                                                        2-methylamino-1,3,4-                                        1,3,4-thiadiazole  thiadiazol-5-yl                                           5-mercapto-2-trifluoromethyl-                                                                    2-trifluoromethyl-1,3,4-                                    1,3,4-thiadiazole  thiadiazol-5-yl                                           2-mercapto-5-p-chlorophenyl-                                                                     5-p-chlorophenyl-1,3,4-                                     1,3,4-thiadiazole  thiadiazol-2-yl                                           3-mercapto-1,2,4-thiadiazole                                                                     1,2,4-thiadiazol-3-yl                                      5-mercapto-1-butyltetrazole                                                                      1-butyltetrazol-5-yl                                       5-mercapto-1-phenyltetrazole                                                                     1-phenyltetrazol-5-yl                                      1-benzyl-1H-tetrazole-5-thiol                                                                    1-benzyl-1H-tetrazol-5-yl                                  5-mercapto-1H-tetrazole                                                                          1H-tetrazol-5-yl                                           5-mercapto-1-p-chlorophenyl-                                                                     1-p-chlorophenyl-1H-                                        1H-tetrazole       tetrazol-5-yl                                             2-mercapto-1,3,4-oxadiazole                                                                      1,3,4-oxadiazol-2-yl                                       2-mercapto-5-phenyl-1,3,4-                                                                       5-phenyl-1,3,4-oxadiazol-                                   oxadiazole         2-yl                                                      2-mercapto-5-benzyl-1,3,4-                                                                       5-benzyl-1,3,4-oxadiazol-                                   oxadiazole         2-yl                                                      5-mercapto-3-phenyl-1,2,4-                                                                       3-phenyl-1,2,4-oxadiazol-                                   oxadiazole         5-yl                                                      2-mercapto-5-ethyl-1,3,4-                                                                        5-ethyl-1,3,4-oxadiazol-                                    oxadiazole         2-yl                                                      2-mercapto-5-trifluoro-                                                                          5-trifluoromethyl-1,3,4-                                    methyl-1,3,4-oxadiazole                                                                          oxadiazol-2-yl                                            1-methyl-5-mercapto-1,2,3-                                                                       1-methyl-1,2,3-triazol-5-yl                                 triazole                                                                     1-ethyl-5-mercapto-1,2,3-                                                                        1-ethyl-1,2,3-triazol-5-yl                                  triazole                                                                     4-methyl-5-mercapto-1,2,3-                                                                       4-methyl-1,2,3-triazol-5-yl                                 triazole                                                                     4-allyl-3-mercapto-1,2,4-                                                                        4-allyl-1,2,4-triazol-3-yl                                  triazole                                                                     4-ethyl-3-mercapto-1,2,4-                                                                        4-ethyl-1,2,4-triazol-3-yl                                  triazole                                                                     3-mercapto-5-methyl-1,2,4-                                                                       5-methyl-1,2,4-triazol-3-yl                                 triazole                                                                     3-mercapto-1,2,4-triazole                                                                        1,2,4-triazol-3-yl                                         4,5-diethyl-3-mercapto-                                                                          4,5-diethyl-1,2,4-triazol-                                  1,2,4-triazole     3-yl                                                      1-cyclopropyl-3-mercapto-                                                                        1-cyclopropyl-1,2,4-                                        1,2,4-triazole     triazol-3-yl                                              3-mercapto-5-methoxymethyl-                                                                      5-methoxymethyl-1,2,4-                                      1,2,4-triazole     triazol-3-yl                                              5-mercapto-3-amino-1,2,4-                                                                        3-amino-1,2,4-triazol-5-yl                                  triazole                                                                     __________________________________________________________________________

EXAMPLE 6

p-Nitrobenzyl 7β-amino-3-(1-methyltetrazol-5-ylthiomethyl)-Δ³-O-2-isocephem-4-carboxylate ##STR135##

Hydrogen sulfide was slowly passed into a solution of the 7β-azido esterof Example 1 (340 mg., 0.72 mmole) and triethylamine (77 mg., 0.76mmole) in 10 ml. of methylene chloride for 50 min. One ml. of 10% HClwas added and the mixture concentrated in vacuo. The residue was mixedwith 25 ml. methylene chloride and washed with 25 ml. each of 10% NaHCO₃and dilute NaCl. The methylene chloride solution was dried, treated withcharcoal, filtered and evaporated in vacuo. The resulting residue wasmixed with methylene chloride, the sulfur filtered off and the solventevaporated in vacuo to give the title product, 276 mg. (86% yield), as ayellow tar. The IR and NMR spectra were consistent with the proposedstructure.

Repeating the above procedure but using the benzyl 7β-azido esterinstead of the p-nitrobenzyl ester gave benzyl7β-amino-3-(1-methyltetrazol-5-ylthiomethyl)-Δ³-O-2-isocephem-4-carboxylate.

EXAMPLE 7

p-Nitrobenzyl 7β-amino-3-(2-methyl-1,3,4-thiadiazol-5-ylthiomethyl)-Δ³-O-2-isocephem-4-carboxylate ##STR136##

The procedure of Example 6 is repeated with the p-nitrobenzyl7β-azido-3-(1-methyltetrazol-5-ylthiomethyl)-Δ³-O-2-isocephem-4-carboxylate used therein replaced by an equimolarweight of p-nitrobenzyl7β-azido-3-(2-methyl-1,3,4-thiadiazol-5-ylthiomethyl)-Δ³-O-2-isocephem-4-carboxylate. The title product is produced as a yellowsolid in 85% yield. The IR and NMR spectra of the product wereconsistent with the proposed structure.

EXAMPLE 8

p-Nitrobenzyl 7β-amino-3-(2-methyl-1,3,4-oxadiazol-5-ylthiomethyl)-Δ³-O-2-isocephem-4-carboxylate ##STR137##

Hydrogen sulfide was bubbled through a solution of the 7β-azido ester ofExample 3 (0.99 g., 2.1 mmole) and trithylamine (0.32 ml., 2.3 mmole) in50 ml. of dichloromethane for 1 min. and the mixture was then stirredfor an additional 45 min. Nitrogen was then bubbled through to removeexcess H₂ S and the solvent was evaporated in vacuo to leave a residueof title product.

EXAMPLE 9

Use of p-nitrobenzyl 7β-azido-3-(1,2,3-triazol-5-ylthiomethyl)-Δ³-O-2-isocephem-4-carboxylate, p-nitrobenzyl7β-azido-3-(1-carboxymethyl-1,2,3,4-tetrazol-5-ylthiomethyl)-Δ.sup.3-O-2-isocephem-4-carboxylate and p-nitrobenzyl7β-azido-3-(1-carboxyethyl-1,2,3,4-tetrazol-5-ylthiomethyl)-Δ.sup.3-O-2-isocephem, respectively, in the procedures of Examples 6-8 givesthe corresponding p-nitrobenzyl 7β-amino-3-(heterocyclic thiomethyl)-Δ³-O-2-isocephem-4-carboxylates.

EXAMPLE 10

Use of the 7β-azido esters of Example 5 in the procedures of Examples6-8 gives the corresponding p-nitrobenzyl 7β-amino-3-(alkyl-, aryl-,aralkyl- or heterocyclicthiomethyl)-Δ³ -O-2-isocephem-4-carboxylates.

EXAMPLE 11

7β-amino-3-(1-methyltetrazol-5-ylthiomethyl)-Δ³ -O-2-isocephem-4-carboxylic acid

Catalytic hydrogenation of the ester of Example 6 with a 20% palladiumhydroxide on carbon catalyst gives the free acid title product.

EXAMPLE 12

Repeating the procedure of Example 11 with the 7β-amino esters ofExamples 7-10 gives the corresponding 7β-amino free acid products.

EXAMPLE 13

Pivaloyloxymethyl 7β-amino-3-(1-methylterazol- 5-ylthiomethyl)-Δ³ -O-2-isocephem-4-carboxylate

The title compound is produced according to the method of Example 2 ofU.K. Specification 1,229,453 by replacing the 7-aminocephalosporanicacid used therein by 7β-amino-3-(1-methyltetrazol5-ylthiomethyl)-Δ³ -O-2-isocephem-4-carboxylic acid.

The respective a(etoxymethyl, methoxymethyl, acetonyl and phenacylesters of 7β-amino-3-(1-methyltetrazol-5-ylthiomethyl)-Δ³ -O-2-isocephem4-carboxylic acid are prepared by substituting in the methodabove for the chloromethyl pivalate used therein an equimolar weight ofchloromethyl acetate, chloromethyl methyl ether, chloroacetone andphenacyl bromide, respectively.

EXAMPLE 14

The procedure of Example 13 is repeated using the 7β-amino carboxylicacids of Example 12 in place of the7β-amino-3-(1-methyltetrazol-5-ylthiomethyl)-Δ³ -O-2-isocephem-4-carboxylic acid used therein. There are produced thecorresponding pivaloyloxymethyl esters. Replacement of chloromethylpivalate by chloromethyl acetate, chloromethyl methyl ether,chloracetone and phenacyl bromide, respectively, produces thecorresponding acetoxymethyl, methoxymethyl, acetonyl and phenacylesters.

EXAMPLE 15

p-Nitrobenzyl7β-(2-thienylacetamido)-3-(1-methyltetrazol-5-ylthiomethyl)-Δ.sup.3 -O-2-isocephem-4-carboxylate ##STR138##

A solution of 2.0 mmole each of p-nitrobenzyl7β-amino-3-(1-methyltetrazol-5-ylthiomethyl)-Δ³ -O-2-isocephem-4-carboxylate, 2-thienyl acetic acid and EEDQ in 20 ml. ofmethylene chloride was stirred at 24° C. for 2 hours. The resultingsolution was washed with 5% NaHCO₃, 10% HCl (2×) and dilute NaCl (20 ml.each), dried and the solvent evaporated in vacuo to give the product asa yellow oil. The product was crystallized from methylene chloride/etherto give the title product in 53% yield; m.p. 165°-168° (dec.). The IRand NMR of the product were consistent with the proposed structure.

EXAMPLE 16

p-Nitrobenzyl7β-(2-thienylacetamido)-3-(2-methyl1,3,4-thiadiazol-5-ylthiomethyl)-.DELTA.³-O- 2-isocephem 4-carboxylate ##STR139##

The procedure of Example 15 was repeated with the 7β-amino ester usedtherein replaced by p-nitrobenzyl7β-amino-3-(2-methyl-1,3,4-thiadiazol5-ylthiomethyl)-Δ³ O-2-isocephem-4-carboxylate. The product was absorbed onto 6 g. of silicagel and placed onto a 24 g. column of silica gel. The column was elutedwith methylene chloride followed by ethyl acetate. The major fractionwas obtained as a yellow foam in 55% yield. Upon recrystallization frommethylene chloride/ether, the title product was obtained; m.p. 127-129°C. The IR and NMR spectra were in agreement with the proposed structure.

Anal. Calc'd. for C₂₄ H₂₁ N₅ O₆ S₃ : C, 49.05; H, 3.60; N, 11.92; S,16.37.Found: C, 47.42; H, 3.54; N, 11.87; S, 16.20.

EXAMPLE 17

p-Nitrobenzyl 7β-phenoxyacetamido-3-(1-methyltetrazol5-ylthiomethyl)-Δ³-O- 2-isocephem-4-carboxylate ##STR140##

A solution of p-nitrobenzyl7β-amino-3-(1-methyltetrazol-5-ylthiomethyl)-Δ³ -O-2-isocephem4-carboxylate (270 mg.), EEDQ (150 mg.) and phenoxyaceticacid (92 mg.) (0.60 mmole each) in 6 ml. of methylene chloride wasmaintained at 24° C. for 2 hours. This solution was diluted to 15 ml.with methylene chloride, washed with 5% NaHCO₃ (25 ml.), 10% HCl (2 × 25ml.) and saturated NaCl (25 ml.), and then dried and absorbed onto 1.7g. of silica gel (grade III). The silica gel was placed on achromatographic column containing 3.4 g. of silica gel (grade III). Thecolumn was eluted with ether changing to ether-methylene chloride (1:1)after fraction 4 (all fractions were 5 ml.). TLC of fractions 8-17showed them to contain one compound (Pf=0.11 on silica gel eluted withether). These fractions were combined and evaporated in vacuo to givethe title product as a yellow tar, 236 mg. (67% yield). The IR and NMRspectra of the product were consistent with the proposed structure.

EXAMPLE 18

Benzyl 7β-phenoxyacetamido-3-(1-methyltetrazol-5ylthiomethyl)-Δ³ -O-2-isocephem-4-carboxylate

The procedure of Example 17 was repeated except that the p-nitrobenzylstarting material was replaced by the corresponding benzyl ester. Thetitle product was obtained upon recrystallization from methylenechloride-ether (1:1); m.p. 86-88° C. U.V. λ_(max) ^(MeOH) 284(ε=10,400). The NMR and IR spectra of the product were consistent withthe proposed structure.

EXAMPLE 19

p-Nitrobenzyl7β-(2-thienylacetamido)-3-(2-methyl1,3,4-oxadiazol-5-yl)-Δ.sup.3 -O-2-isocephem-4-carboxylate ##STR141##

The p-nitrobenzyl ester residue of Example 8 was dissolved in 50 ml. ofdichloromethane and treated with 2-thienylacetic acid (0.31 g., 2.2mmoles) followed by EEDQ (0.54 g., 2.2 mmoles). The mixture was stirredat room temperature for 16 hours and then washed with water, 1% NaHCO₃,water, 10% HCl, water and brine. It was then dried over sodium sulfateand evaporated in vacuo. The residue was purified by dry-columnchromatography on silica gel (50 g., activity III) eluting with 25%ethyl acetate in ether to give 0.61 g. of pure title product in 50%yield. The NMR of the product was in agreement with the proposedstructure.

EXAMPLE 20

7β(2-thienylacetamido)-3-(1-methyletrazol-5-ylthiomethyl)-Δ.sup.3 -O-2-isocephem-4-carboxylic acid 20

The p-nitrobenzyl ester from Example 15 (1.00 mmole) in 50 ml. of ethylacetate and 25 ml. of n-butanol to which 10 ml. of 0.1 M HCl and 500 mg.of 20% palladium hydroxide-on-diatomaceous-earth had been added washydrogenated on a Parr apparatus at 24° C. and 50 p.s.i. for 3 hours.The catalyst was filtered off and the solution extracted with 1% NaHCO₃(containing some NaCL) (3 × 25 ml.). The aqueous extract was cooled to0° C., acidified to pH 1 with 10% HCl and saturated with NaCl. Theacidified aqueous was extracted with methylene chloride (4 × 50 ml.).The methylene chloride was concentrated in vacuo to 100 ml. and afterstanding, the title product was collected as a colorless solid in 18.5%yield; m.p. 182- 184° (dec.). U.V. λ _(max) ^(THF) 280 (ε=8500). The IRspectrum was consistent with the proposed structure.

Anal. Calc'd. for C₁₆ H₁₆ N₆ O₅ S₂ : C, 44.03; H, 3.70; N, 19.26. Found:C, 44.10, H, 3.71; N, 19.07.

A sample of the title product (called BC-L70) after solution in waterand dilution with Nutrient Broth was found to exhibit the followingMinimum Inhibitory Concentrations (M.I.C.) in mcg./ml. versus theindicated microorganisms as determined by overnight incubation at 37° C.by tube dilution.

    ______________________________________                                        M.I.C. in mcg./ml.                                                                                        Cepha-  Cepha-                                    Organism           BC-L70   lexin   lothin                                    ______________________________________                                        D. pneumoniae A9585    .016     .25   .13                                     +5% serum*                                                                    Str. pyrogenes                                                                              A9604    .016     .25   .06                                     +5% serum*                                                                    S. aureus Smith.sup.+                                                                       A9537    .13       1    .13                                     S. aureus Smith.sup.+                                                                       A9537     1        1     .5                                     +50% serum                                                                    S. aureus BX1633-2                                                                          A9606     .5       2    .25                                     at 10.sup.-dil'n                                                              S. aureus BX1633-2                                                                          A9606     2        4     .5                                     at 10.sup.-.sup.2 dil'n                                                       S. aureus meth.-                                                                            A15097    1        8     1                                      resist.; at 10.sup.-.sup.3                                                    dil'n                                                                         Sal. enteritidis.sup.+                                                                      A9531    .03       4     .5                                     E. coli Juhl.sup.+                                                                          A15119    1        8     16                                     E. coli.sup.+ A9675     16       8     63                                     K. pneumoniae.sup.+                                                                         A9977    .25       4     4                                      K. pneumoniae.sup.+                                                                         A15130    8        16    16                                     Pr. mirabilis.sup.+                                                                         A9900    .13       4     1                                      Pr. morganii.sup.+                                                                          A15153    32      125   125                                     Ps. aeruginosa.sup.+                                                                        A9843A   125      125   125                                     Ser. marcescen.sup.+                                                                        A20019   125      125   125                                     Ent. cloacae  A9656    125      125   125                                     Ent. cloacae  A9657     1        4     4                                      Ent. cloacae  A9659    125      125   125                                     ______________________________________                                         *50% Nutrient Broth - 45% Antibiotic Assay Broth at 10.sup.-.sup.4            dilution.                                                                

EXAMPLE 21

7β-(2-thienylacetamido)-3-(2-methyl-1,3,4-thiadiazol5-ylthiomethyl)-.DELTA.³-O- 2-isocephem-4-carboxylic acid

The procedure of Example 20 was repeated up to the point of preparingthe acidified aqueous extract with the p-nitrobenzyl ester startingmaterial replaced by p-nitrobenzyl7β-(2-thienylacetamido)-3-(2-methyl-1,3,4-thiadiazol-5-ylthiomethyl)-Δ³-O- 2-isocephem-4-carboxylate. The acidified aqueous was extracted withtetrahydrofuran/ethyl acetate (9:1) (2 × 50 ml.).

The organic extract was washed with saturated NaCl (50 ml.), dried,treated with charcoal and the solvent evaporated in vacuo to give abrown oil. The oil was crystallized from ethyl acetate/ether to give thetitle product in 39% yield; m.p. 100°-105° C. (dec.). U.V. λ_(max)^(THF/H) ₂ O 276 (ε=12,500). The IR and NMR were consistent with theproposed structure.

Anal. Calc'd. for C₁₇ H₁₆ N₄ O₅ S₃. 1/2 CH₃ CO₂ C₂ H₅ : C, 45.96; H,4.06; N, 11.29; S, 19.37. Found: C, 45.69; H, 3.97; N, 11.30; S, 19.63.

M.I.C. data for the product (called BC-L71) is shown in the followingtable.

    ______________________________________                                        M.I.C. in mcg./ml.                                                                                       Cepha-   Cepha-                                    Organism          BC-L71   lexin    lothin                                    ______________________________________                                        D. pneumoniae                                                                              A9585    .016      .5    .06                                      +5% serum*           .008      .5    .06                                     Str. pyogenes                                                                              A9604    .016     .25    .06                                      +5% serum*           .008     .13    .06                                     S. aureus Smith.sup.+                                                                      A9537     .25      1     .13                                                            .06      .5    .06                                     S. aureus Smith.sup.+                                                                      A9537     2        2      .5                                      +50% serum            2        2      .5                                     S. aureus BX1633-2                                                                         A9606     .5       2     .25                                      at 10.sup.-.sup.3 dil'n                                                                             .13      1     .13                                     S. aureus BX1633-2                                                                         A9606     4        4      .5                                      at 10.sup.-.sup.2 dil'n                                                                             2        2     .25                                     S. aureus meth.-                                                                           A15097    4        16     1                                       resist.; at 10.sup.-.sup.3                                                                          2        16     1                                       dil'n                                                                        Sal. enteritidis.sup.+                                                                     A9531     .06      2     .13                                                           .016      2     .25                                     E. coli Juhl .sup.+                                                                        A15119    8        8      8                                                             4        8      8                                      E. coli .sup.+                                                                             A9675     32       16     32                                                            32       16     63                                     K. pneumoniae .sup.+                                                                       A9977     1        4      2                                                             .5       4      1                                      K. pneumoniae .sup.+                                                                       A15130    32       16     16                                                            32       16     16                                     Pr. mirabilis .sup.+                                                                       A9900     1        8      1                                                             .5       4      1                                      Pr. morganii .sup.+                                                                        A15153   125      125    125                                                            32      125    125                                     Ps. aeruginosa .sup.+                                                                      A9843A   125      125    125                                                           125      125    125                                     Ser. marcescens .sup.+                                                                     A20019   125      125    125                                                           125      125    125                                     Ent. cloacae A9656    125      125    125                                     Ent. cloacae A9657     4        4      4                                                             16       4      8                                      Ent. cloacae A9659    125      125    125                                                           125      125    125                                     ______________________________________                                         *50% Nutrient Broth - 45% Antibiotic Assay Broth at 10.sup.-.sup.4            dilution.                                                                

EXAMPLE 22

7β-(2-thienylacetamido)-3-(2-methyl-1,3,4-oxadiazol5-ylthiomethyl)-.DELTA.³-O- 2-isocephem-4-carboxylic acid

The p-nitrobenzyl ester of Example 19 (0.60 g., 0.15 mmole) wasdissolved in 125 ml. of ethyl acetate and 40 ml. of n-butanol. Thesolution was treated with 1.05 ml. of IN HCl (1.05 mmole) andhydrogenated over 0.60 g. of 20% palladium hydroxide on carbon for 31/2hours at 50 p.s.i.g. The mixture was then filtered, washing well withethyl acetate, and evaporated in vacuo. The residue was slurried withether and extracted with 1% NaHCO₃ (2 × 50 ml.) followed by 25 ml. ofwater. The aqueous extracts were acidified in the cold with 10% HCl andextracted with ethyl acetate (3 × 30 ml.). The solution was dried oversodium sulfate and evaporated in vacuo. The solid residue was slurriedwith ether, filtered and dried giving 69 mg. (15% yield) of the titleproduct; m.p. 170- 173°. U.V. λ _(max) ^(THF=) 278 nm (ε=12,453).

Anal. Calc'd. for C₁₇ H₁₆ N₄ O₆ S₂ : C, 46.78; H, 3.70; N, 12.84. Found:C, 43.95; H, 3.57; N, 11.35; residue: 3.05.

M.I.C. data for the product (called BC-L76) is shown in the followingtable.

    ______________________________________                                        M.I.C. in mcg./ml.                                                                                       Cepha-   Cepha-                                    Organism          BC-L76   lexin    lothin                                    ______________________________________                                        D. pneumoniae                                                                              A9585    .016      .25   .016                                     +5% serum*                                                                   Str. pyogenes                                                                              A9604    .008      .13    .03                                     +5% serum*                                                                   S. aureus Smith .sup.+                                                                     A9537     .06       .5    .06                                    S. aureus Smith .sup.+                                                                     A9537      1        1     .25                                     +50% serum                                                                   S. aureus BX1633-2                                                                         A9606     .5        2     .13                                     at 10.sup.-.sup.3 dil'n                                                      S. aureus BX1633-2                                                                         A9606     .5        4     .13                                     at 10.sup.-.sup.2 dil'n                                                      S. aureus meth.-                                                                           A15097     2        32     2                                      resist.; at 10.sup.-.sup.3                                                    dil'n                                                                        Sal. enteritidis .sup.+                                                                    A9531     .5        2     .13                                    E. coli Juhl .sup.+                                                                        A15119     8        8     16                                     E. coli .sup.+                                                                             A9675     63        16    63                                     K. pneumoniae .sup.+                                                                       A9977      1        4      2                                     K. pneumoniae .sup.+                                                                       A15130    63        8     32                                     Pr. mirabilis .sup.+                                                                       A9900     .5        4     .5                                     Pr. morganii .sup.+                                                                        A15153    125     >125   >125                                    Ps. aeruginosa .sup.+                                                                      A9843A   >125     >125   >125                                    Ser. marcescens .sup.+                                                                     A20019   >125     >125   >125                                    Ent. cloacae A9656    >125     >125   >125                                    Ent. cloacae A9657     16        4      8                                     Ent. cloacae A9659    >125     >125   >125                                    ______________________________________                                         *50% Nutrient Broth - 45% Antibiotic Assay Broth at 10.sup.-.sup.4            dilution.                                                                

EXAMPLE 23

7β-phenoxyacetamido-3-(1-methyltetrazol-5-ylthiomethyl)-Δ³ -O-2-isocephem-4-carboxylic acid

A solution of the p-nitrobenzyl ester prepared according to Example 17(235 mg., 0.405 mmole) in 12 ml. of tetrahydrofuran (peroxide free) and2.4 ml. of ethanol with 140 mg. of 10% palladium-oncharcoal washydrogenated (Parr shaker) at 24° and 50 p.s.i. for 4 hours. Thecatalyst was filtered off and the solvent evaporated in vacuo. Theresidue was dissolved in 25 ml. of ethyl acetate and washed with 25 ml.each of 10% HCl and water. The acqueous layers were combined andextracted with 10 ml. of ethyl acetate (which was washed with 10 ml. ofwater). The combined ethyl acetate was extracted with 1% NaHCO₃ (2 × 10ml.). The combined bicarbonate extracts were acidified with 10% HCl andextracted with ethyl acetate. The ethyl acetate extract was washed withwater and saturated NaCl and then dried and evaporated in vacuo to give67 mg. (37% yield) of crude title product. The product was crystallizedfrom chloroform to give title product with m.p. 136- 138° (dec.). U.V. λ_(max) ^(MeOH) 276 (ε=10,600).

Anal. Calc'd. for C₁₈ H₁₈ N₆ O₆ S: C, 48.43; H, 4.06; N, 18.83; S, 7.18.Found: C, 46.70; H, 3.94 N, 17.74; S, 6.98; Residue: 1.16.

M.I.C. data for the product (called BC-L62) is shown in the followingtable.

    ______________________________________                                        M.I.C. in mcg./ml.                                                                                       Cepha-   Cepha-                                    Organism          BC-L62   lexin    lothin                                    ______________________________________                                        D. pneumoniae                                                                              A9585     .13      .13    .03                                     +5% serum*                                                                   Str. pyogenes                                                                              A9604     .13      .06    .03                                     +5% serum*                                                                   S. aureus Smith .sup.+                                                                     A9537     .25      .25    .06                                    S. aureus Smith .sup.+                                                                     A9537      1        1     .25                                     +50% serum                                                                   S. aureus BX1633-2                                                                         A9606      2        2     .13                                     at 10.sup.-.sup.3 dil'n                                                      S. aureus BX1633-2                                                                         A9606     16        4     .25                                     at 10.sup.-.sup.2 dil'n                                                      S. aureus meth.-                                                                           A15097     4       16      1                                      resist.; at 10.sup.-.sup.3                                                    dil'n                                                                        Sal. enteritidis .sup.+                                                                    A9531     .5        4     .13                                    E. coli Juhl .sup.+                                                                        A15119     4        8     16                                     E. coli .sup.+                                                                             A9675     32       16     32                                     K. pneumoniae .sup.+                                                                       A9977       2       4      1                                     K. pneumoniae .sup.+                                                                       A15130    32       16     32                                     Pr. mirabilis .sup.+                                                                       A9900      2        4     .5                                     Pr. morganii .sup.+                                                                        A15153    63       125   >125                                    Ps. aeruginosa .sup.+                                                                      A9843A   >125     >125   >125                                    Ser. marcescens .sup.+                                                                     A20019   >125     >125   >125                                    Ent. cloacae A9656    >125     >125   >125                                    Ent. cloacae A9657     32        4      4                                     Ent. cloacae A9659    125      >125   >125                                    ______________________________________                                         *50% Nutrient Broth - 45% Antibiotic Assay Broth at 10.sup.-.sup.4            dilution.                                                                

EXAMPLE 24

7β-[d-α-amino-α-phenylacetamido]-3-(1-methyltetrazol-5-ylthiomethyl)-Δ³-0-2-isocephem-4-carboxylic acid (hydrochloride salt) ##STR142##

A mixture containing p-nitrobenzyl7β-amino-3-(1-methyltetrazol-5-ylthiomethyl)-Δ³-0-2-isocephem4-carboxylate (2.8 g., 6.25 mmole), EEDQ (1.55 g., 6.25mmole) and D(-) α-azidophenyl acetic acid (1.11 g., 6.25 mmole) in 200ml. methylene chloride was kept at room temperature (protected frommoisture by a calcium chloride drying tube) for 16 hours. It was thenwashed successively with 10% HCl, water, 5% NaHCO₃ and brine, dried (Na₂SO₄) and evaporated in vacuo to dryness leaving a yellow amorphoussolid. This was suspended in ether and filtered to give 3.67 g. of asolid which was identified by IR and NMR as p-nitrobenzyl7β-[α-azidoα-phenylacetamido]-3-(1-methyltetrazol-5-ylthiomethyl)Δ³-0-2-isocephem-4-carboxylate; m.p. 132-134° C.

Anal. Calc'd. for C₂₅ H₂₂ N₁₀ O₇ S: C, 49.50; H, 3.65; N, 23.09. Found:C, 49.47; H, 3.64; N, 22.86.

A mixture consisting of the above ester (0.40 g., 0.65 mmole) and 20%Pd(OH)₂ on Celite (0.50 g.) in 50 ml. ethyl acetate, 50 ml. n-butanoland 6.5 ml. of 0.1N HCl was shaken on a Parr apparatus under 60 p.s.i.of hydrogen for 6 hours. The solid was filtered off on a Celite padwhich was washed well with absolute ethanol. The combined filtrates wereevaporated in vacuo at 40° C. to leave a yellow amorphous solid. Thiswas triturated with ether and collected by filtration to give 0.21 g.solid; m.p. 110-120° (decomp.). U.V. λ_(max) ^(THF) =274, ε_(max) =7550.The IR and NMR spectra of the product were in agreement with theproposed structure.

M.I.C. data for the product (called BC-L78) is shown in the followingtable. Purity of the sample was estimated to be about 25%.

    __________________________________________________________________________    M.I.C. in mcg./ml.                                                            Organism           BC-L78                                                                             Cephalexin                                                                          Cephalothin                                     __________________________________________________________________________    D. pneumoniae                                                                            A9585   0.25 0.25  .03                                              +5% serum*                                                                   Str. pyogenes                                                                            A9604   0.13 0.13  .03                                              +5% serum*                                                                   S. aureus Smith .sup.+                                                                   A9537   2    0.5   .06                                             S. aureus Smith .sup.+                                                                   A9537   8    1     0.5                                              +50% serum                                                                   S. aureus BX1633-2                                                                       A9606   8    2     0.25                                             at 10.sup.-.sup.3 dil'n                                                      S. aureus BX1633-2                                                                       A9606   63   8     0.5                                              at 10.sup.-.sup.2 dil'n                                                      S. aureus meth.-                                                                         A15097 (37°)                                                                   63   32    1                                                resist.; at 10.sup.-.sup.3                                                                   (28°)                                                                     >125 125   63                                               dil'n                                                                        Sal. enteritidis .sup.+                                                                  A9531   0.5  2     0.25                                            E. coli Juhl .sup.+                                                                      A15119  2    8     16                                              E. coli .sup.+                                                                           A9675   16   16    63                                              K. pneumoniae .sup.+                                                                     A9977   1    4     1                                               K. pneumoniae .sup.+                                                                     A15130  8    16    16                                              Pr. mirabilis .sup.+                                                                     A9900   1    4     1                                               Pr. morganii .sup.+                                                                      A15153  32   >125  >125                                            Ps. aeruginosa .sup.+                                                                    A9843A  >125 >125  >125                                            Ser. marcescens .sup.+                                                                   A20019  >125 >125  >125                                            Ent. cloacae                                                                             A9656   >125 >125  >125                                            Ent. cloacae                                                                             A9657   4    4     4                                               Ent. cloacae                                                                             A9659   16   >125  >125                                            __________________________________________________________________________     *50% Nutrient Broth - 45% Antibiotic Assay Broth at 10.sup.-.sup.4            dilution.                                                                

EXAMPLE 25

Repeating the general N-acylation procedures of Examples 15, 16, 17, 18,19, or 24 to react the following acylating agents with7β-amino-3-(1-methyltetrazol-5ylthiomethyl)-Δ³-0-2-isocephem-4-carboxylic acid (or an ester or salt thereof), thefollowing products are obtained after removal of any functional blockinggroups.

    __________________________________________________________________________    Acylating Agent    Product                                                    __________________________________________________________________________    mixed anhydride of                                                                           7β-(2-Aminomethylphenyl-                                  potassium 2-(1-carbo-                                                                        acetamido)-3-(1-methyltetrazol-                                methoxypropen-2-ylamino-                                                                     5-ylthiomethyl)-Δ.sup.3 -0-2-                            methyl)phenylacetate with                                                                    isocephem-4-carboxylic acid                                    isobutyl chloroformate                                                        α-benzoylureidophenyl-                                                                 7β-(α-Benzoylureidophenyl-                          acetic acid    acetamido)-3-(1-methyltetrazol-                                               5-ylthiomethyl)-Δ.sup.3 -0-2-                                           isocephem-4-carboxylic acid                                    2,6-dimethoxybenzoyl                                                                         7β-(2,6-Dimethoxybenzamido)-                              chloride       3-(1-methltetrazol-5-ylthio-                                                  methyl)-Δ.sup.3 -0-2-isocephem-4-                                       carboxylic acid                                                D-anhydro-o-carboxy-                                                                         7β-(D-α-Hydroxyphenylacetamido)-                    mandelic acid  3-(1-methyltetrazol-5-ylthio-                                                 methyl)-Δ.sup.3 -0-2-isocephem-4-                                       carboxylic acid                                                3 benzyl-1,2,4-oxadiazole-                                                                   7β-[N-(Phenylacetimidoyl)-                                5-one-4-acetic acid                                                                          aminoacetamido]-3-(1-methylte-                                                trazol-5-ylthiomethyl)-Δ.sup.3 -0-2-                                    isocephem-4-carboxylic acid                                    valeric acid   7β-Valeramido-3-(1-methylte-                                             trazol-5-ylthiomethyl)-Δ.sup.3 -0-2-                                    isocephem-4-carboxylic acid                                    phenylacetic acid                                                                            7β-Phenylacetamido-3-(1-                                                 methyltetrazol-5-ylthiomethyl)-                                               Δ.sup.3 -0-2-isocephem-4-carboxylic                                     acid                                                           3-thienylacetyl chloride                                                                     7β-(3-Thienylacetamido)-3-                                               (1-methyltetrazol-5-ylthio-                                                   methyl)-Δ.sup.3 -0-2-isocephem-4-                                       carboxylic acid                                                α-carboxybenzyl-phenyl-                                                                7β-[α-carboxy-α-phenyl-                       acetic acid    acetamido]-3-(1-methyltetrazol-                                               5-ylthiomethyl)-Δ.sup.3 -0-2-isocephem-                                 4-carboxylic acid                                              o-hydroxyphenylacetic                                                                        7β-(o-Hydroxyphenylacetamido)-                            acid           3-(1-methyltetrazol-5-ylthiomethyl)-                                          Δ.sup.3 -0-2-isocephem-4-carboxylic                                     acid                                                           cyanoacetic acid                                                                             7β-[cyanoacetamido)-3-(1-                                                methyltetrazol-5-ylthiomethyl)-                                               Δ.sup.3 -0-2-isocephem-4-carboxylic                                     acid                                                           α-cyanopropionic acid                                                                  7β-(α-cyanopropionamido)-3-                                        (1-methyltetrazol-5-ylthio-                                                   methyl)-Δ.sup.3 -0-2-isocephem-4-                                       carboxylic acid                                                2-(2H)-tetrazoleacetic                                                                       7β-[2-(2H)-tetrazolylacetamido]-                          acid           3-(1-methyltetrazol-5-ylthiomethyl)-                                          Δ.sup.3 -0-2-isocephem-4-carboxylic acid                 3-(o-chlorophenyl)-5-                                                                        7β-[3-(o-chlorophenyl)-5-                                 methyl-4-isoxazole-                                                                          methylisoxazol-4-ylcarboxamido]-                               carboxylic acid chloride                                                                     3-(1-methyltetrazol-5-ylthio-                                                 methyl)-Δ.sup.3 -0-2-isocephem-4-                                       carboxylic acid                                                1-(1H)-tetrazolylacetyl                                                                      7β-[1-(1H)-tetrazolylacetamido]-                          chloride       3-(1-methyltetrazol-5-ylthio-                                                 methyl)-Δ.sup.3 -0-2-isocephem-4-                                       carboxylic acid                                                __________________________________________________________________________

EXAMPLE 26

Repeating the general N-acylation procedures of the examples above toreact the following acylating agents with7β-amino-3-(1-methyltetrazol-5-ylthiomethyl)-Δ³-0-2-isocephem-4-carboxylic acid (or an ester or salt thereof), thefollowing products are obtained after removal of any functional blockinggroups.

    __________________________________________________________________________    Acylating Agents   Product                                                    __________________________________________________________________________    4-nitrophenylacetyl                                                                           7β-(4-nitrophenylacetamido)-                             chloride        3-(1-methyltetrazol-5-ylthio-                                                 methyl)-Δ.sup.3 -0-2-isocephem-4-                                       carboxylic acid                                               p-fluorophenylacetyl                                                                          7β-(p-fluorophenylacetamido)-                            chloride        3-(1-methyltetrazol-5-ylthio-                                                 methyl)-Δ.sup.3 -0-2-isocephem-4-                                       carboxylic acid                                               p-acetoxyphenylacetyl                                                                         7β-(p-acetoxyphenylacetamido)-                           chloride        3-(1-methyltetrazol-5-ylthio-                                                 methyl)-Δ.sup.3 -0-2-isocephem-4-                                       carboxylic acid                                               o-chlorophenylacetyl                                                                          7β-(o-chlorophenylacetamido)-                            chloride        3-(1-methyltetrazol-5-ylthio-                                                 methyl)-Δ.sup.3 -0-2-isocephem-4-                                       carboxylic acid                                               p-aminophenylacetyl                                                                           7β-(p-aminophenylacetamido)-                             chloride        3-(1-methyltetrazol-5-ylthio-                                                 methyl)-Δ.sup.3 -0-2-isocephem-4-                                       carboxylic acid                                               p-methylphenylacetyl                                                                          7β-(p-methylphenylacetamido)-                            chloride        3-(1-methyltetrazol-5-ylthio-                                                 methyl)-Δ.sup.3 -0-2-isocephem-4-                                       carboxylic acid                                               4-guanidinophenylacetyl                                                                       7β-(4-guanidinophenylacetamido)-                         chloride hydrochloride                                                                        3-(1-methyltetrazol-5-ylthio-                                                 methyl-Δ.sup.3 -0-2-isocephem-4-                                        carboxylic acid                                               4-isopropylphenylacetyl                                                                       7β-(4-isopropylphenylacetamido)-                         chloride        3-1(1-methyltetrazol-5-ylthio-                                                methyl)-Δ.sup.3 -0-2-isocephem-4-                                       carboxylic acid                                               4-methylthiophenylacetyl                                                                      7β-(4-methylthiophenylacetamido)-                        chloride        3-(1-methyltetrazol-5-ylthiomethyl)-                                          Δ.sup.3 -0-2-isocephem-4-carboxylic                                     acid                                                          4-cyanophenylacetyl                                                                           7β-(4-cyanophenylacetamido)-                             chloride        3-(1-methyltetrazol-5-ylthiomethyl)-                                          Δ.sup.3 -0-2-isocephem-4-carboxylic                                     acid                                                          4-methoxyphenylacetyl                                                                         7β-(4-methoxyphenylacetamido)-                           chloride        3-(1-methyltetrazol-5-ylthio-                                                 methyl)-Δ.sup.3 -0-2-isocephem-4-                                       carboxylic acid                                               2,6-dimethoxyphenylacetyl                                                                     7β-(2,6-dimethoxyphenylacetamido)-                       chloride        3-(1:methyltetrazol-5-ylthio-                                                 methyl)-Δ .sup.3 -0-2-isocephem-4-                                      carboxylic acid                                               3-sulfamylphenylacetyl                                                                        7β-(3-sulfamylphenylacetamido)-                          chloride        3-(1-methyltetrazol-5-ylthio-                                                 methyl)-Δ.sup.3 -0-2-isocephem-4-                                       carboxylic acid                                               2-methyl-4-chlorophenyl-                                                                      7β-(2-methyl-4-chlorophenyl-                             acetyl chloride acetamido)-3-(1-methyltetrazo-                                                5-ylthiomethyl)-Δ.sup.3 -0-2-isocephem-                                 4-carboxylic acid                                             sydnone-3-acetyl chloride                                                                     7β-(sydnone-3-acetamido)-3-                                              (1-methyltetrazol-5-ylthio-                                                   methyl)-Δ.sup.3 -0-2-isocephem-4-                                       carboxylic acid                                               sydnone-4-acetyl chloride                                                                     7β-(sydnone-4-acetamido)-3-                                              (1-methyltetrazol-5-ylthio-                                                   methyl)-Δ.sup.3 -0-2-isocephem-4-                                       carboxylic acid                                               2-furylacetyl chloride                                                                        7β-(2-furylacetamido)-3-                                                 (1-methyltetrazol-5-ylthio-                                                   methyl)-Δ.sup.3 -0-2-isocephem-4-                                       carboxylic acid                                               3-furylacetyl chloride                                                                        7β-(3-furylacetamido)-3-                                                 (1-methyltetrazol-5-ylthio-                                                   methyl)-Δ.sup.3 -0-2-isocephem-4-                                       carboxylic acid                                               1,2,5-thiadiazole-3-                                                                          7β-(1,2,5-thiadiazole-3-                                 acetyl chloride acetamido)-3-(1-methyltetrazol-                                               5-ylthiomethyl)-Δ.sup.3 -0-2-isocephem-                                 4-carboxylic acid                                             1-cyclohexenylacetyl                                                                          7β-(1-cyclohexenylacetamido)-                            chloride        3-(1-methyltetrazol-5-ylthio-                                                 methyl)-Δ.sup.3 -0-2-isocephem-4-                                       carboxylic acid                                               1,4-cyclohexadienyl-                                                                          7β-(1,4-cyclohexadienyl-                                 acetyl chloride acetamido)-3-(1-methyltetrazol-                                               5-ylthiomethyl)-Δ.sup.3 -0-2-isocephem-                                 4-carboxylic acid                                             3-(1,4-cyclohexadien-                                                                         7β-[3-(1,4-cyclohexadien-1-yl)-                          1-yl)propionyl chloride                                                                       propionamido]-3-(1-methyltetrazol-                                            5-ylthiomethyl)-Δ.sup.3 -0-2-isocephem-                                 4-carboxylic acid                                             isothiazol-4-yl-acetic                                                                        7β-(isothiazol-4-yl-acetamido)-                          acid            3-(1-methyltetrazol-5-ylthio-                                                 methyl)-Δ.sup.3 -0-2-isocephem-4-                                       carboxylic acid                                               isothiazol-5-ylacetic                                                                         7β-(isothiazol-5-yl-acetamido)-                          acid            3-(1-methyltetrazol-5-ylthio-                                                 methyl)-Δ.sup.3 -0-2-isocephem-4-                                       carboxylic acid                                               isothiazol-3-yl-acetic                                                                        7β-(isothiazol-3-yl-acetamido)-                          acid            3-(1-methyltetrazol-5-ylthio-                                                 methyl)-Δ.sup.3 -0-2-isocephem-4-                                       carboxylic acid                                               5-phenyl-1,3,4-theiadizolyl-                                                                  7β-(5-phenyl-1,3,4-thiadiazol-                           2-yl-acetyl chloride                                                                          2-yl-acetamido)-3-(1-methyltetra-                                             zol-5-ylthiomethyl)-Δ.sup.3 -0-2-                                       isocephem-4-carboxylic acid                                   thiazol-2-yl-acetyl                                                                           7β-(thiazol-2-yl-acetamido)-                             chloride        3-(1-methyltetrazol-5-ylthio-                                                 methyl)-Δ.sup.3 -0-2-isocephem-4-                                       carboxylic acid                                               imidazol-2-yl-acetyl                                                                          7β-(imidazol-2-yl-acetamido)-                            chloride        3-(1:methyltetrazol-5-ylthio-                                                 methyl)-Δ.sup.3 -0-2-isocephem-4-                                       carboxylic acid                                               1,2,3-triazol-4-yl-                                                                           7β-(1,2,3-triazol-4-yl-acetamido)-                       acetic acid     3-(1-methyltetrazol-5-ylthio-                                                 methyl)-Δ.sup.3 -0-2-isocephem-4-                                       carboxylic acid                                               oxazol-2-yl-acetyl                                                                            7β-(oxazol-2-yl-acetamido)-3-                            chloride        (1-methyltetrazol-5-ylthiomethyl)-                                            Δ.sup.3 -0-2-isocephem-4-carboxylic                                     acid                                                          4-pyridylacetyl 7β-(4-pyridylacetamido)-3-                               chloride        (1-methyltetrazol-5-ylthiomethyl)-                                            Δ.sup.3 -0-2-isocephem-4-carboxylic                                     acid                                                          3-pyridylacetyl chloride                                                                      7β-(3-pyridylacetamido)-3-                                               (1-methyltetrazol-5-ylthiomethyl)-                                            Δ.sup.3 -0-2-isocephem-4-carboxylic                                     acid                                                          3-phenylpropionyl chloride                                                                    7β-(3-phenylpropionamido)-3-                                             (1-methyltetrazol-5-ylthiomethyl)-                                            Δ.sup.3 -0-2-isocephem-4-carboxylic                                     acid                                                          3-(p-chlorophenyl )propionyl                                                                  7β-[3-(p-chlorophenyl)propion-                           chloride        amido)-3-(1-methyltetrazol-5-                                                 ylthiomethyl)-Δ.sup.3 -0-2-isocephem-                                   4-carboxylic acid                                             3-(p-methoxyphenyl)propionyl                                                                  7β-[p-methoxyphenyl)propion-                             chloride        amido)-3-(1:methyltetrazol-5-                                                 ylthiomethyl)-Δ.sup.3 -0-2-isocephem-                                   4-carboxylic acid                                             3-(p-sulfamylphenyl)-                                                                         7β-[3-(p-sulfamylphenyl)propion-                         propionyl chloride                                                                            amido]-3-(1-methyltetrazol-3-                                                 ylthiomethyl)-Δ.sup.3 -0-2-isocephem-                                   4-carboxylic acid                                             3-(3,4-dimethoxyphenyl)-                                                                      7β-[3-(3,4-dimethoxyphenyl)-                             propionyl chloride                                                                            propionamido]-3-(1-methyltetrazol-                                            5-ylthiomethyl)-Δ.sup.3 -0-2-isocephem-                                 4-carboxylic acid                                             3-(p-hydroxyphenyl)-                                                                          7β-[3-(p-hydroxyphenyl)propion-                          propionic acid  amido]-3-(1-methyltetrazol-5-                                                 ylthiomethyl)-Δ.sup.3 -0-2-isocephem-                                   4-carboxylic acid                                             3-(p-nitrophenyl)-                                                                            7β-[3-(p-nitrophenyl)propion-                            propionic acid  amido]-3-(1-methyltetrazol-5-                                                 ylthiomethyl)-Δ.sup.3 -0-2-isocephem-                                   4-carboxylic acid                                             3-(2-thienyl)propionyl                                                                        7β-[3-(2-thienyl)propionamido]-                                          3-(1-methyltetrazol-5-ylthio-                                                 methyl)-Δ.sup.3 -0-2-isocephem-4-                                       carboxylic acid                                               3-(3-thienyl)propionyl                                                                        7β-[3-(3-thienyl)propionamido]-                                          3-(1-methyltetrazol-5-ylthio-                                                 methyl)-Δ.sup.3 -0-2-isocephem-4-                                       carboxylic acid                                               cyclohexylacetic acid                                                                         7β-(cyclohexylacetamido)-3-                                              (1-methyltetrazol-5-ylthiomethyl)-                                            Δ.sup.3 -0-2-isocephem-4-carboxylic acid                3-phenyl-5-methylisoxazol-                                                                    7β-(3-phenyl-5-methylisoxazol-                           4-yl-acetic acid                                                                              4-yl-acetamino)-3-(1-methyltetrazol-                                          5-ylthiomethyl)-Δ.sup.3 -0-2-isocephem-                                 4-carboxylic acid                                             o-aminomethylphenylacetic                                                                     7β-(o-aminomethylphenyl-                                 acid            acetmido)-3-(1-methyltetrazol-                                                5-ylthiomethyl)-Δ.sup.3 -0-2-isocephem-                                 4-carboxylic acid                                             3-methoxy-4-furazanacetyl                                                                     7β-(3-methoxy-4-furazan-                                 chloride        acetamido)-3-(1-methyltetrazol-                                               5-ylthiomethyl)-Δ.sup.3 -0-2-isocephem-                                 4-carboxylic acid                                             __________________________________________________________________________

EXAMPLE 27

Repeating the general N-acylation procedures of the examples above toreact the following acylating agents with7β-amino-3-(1-methyltetrazol-5-ylthiomethyl)Δ³-0-2-isocephem-4-carboxylic acid (or an ester or salt thereof), thefollowing products are obtained after removal of any functional blockinggroups.

    __________________________________________________________________________    Acylating Agent    Product                                                    __________________________________________________________________________    p-nitrophenoxyacetic                                                                       7β-(p-nitrophenoxyacetamido-                                acid         3-(1-methyltetrazol-5-ylthiomethyl)-                                          Δ.sup.3 -O-2-isocephem-4-carboxylic acid                   p-fluorophenoxyacetic                                                                      7β-(p-fluorophenoxyacetamido)-                              acid         3-(1-methyltetrazol-5-ylthiomethyl)-                                          Δ.sup.3 -O-2-isocephem-4-carboxylic acid                   o-chlorophenoxyacetic                                                                      7β-(o-chlorophenoxyacetamido)-                              acid         3-(1-methyltetrazol-5-ylthio-                                                 methyl)-Δ.sup.3 -0-2-isocephem-4-                                       carboxylic acid                                                  p-sulfamylphenoxyacetic                                                                    7β-(p-sulfamylphenoxyacetamido)-                            acid         3-(1-methylmetrazol-5-ylthio-                                                 methyl)-Δ.sup.3 -0-2-isocephem-4-                                       carboxylic acid                                                  p-methylphenoxyacetic                                                                      7β-(p-methylphenoxyacetamido)-                              acid         3-(1-methyltetrazol-5-ylthio-                                                 methyl)-Δ.sup.3 -0-2-isocephem-4-                                       carboxylic acid                                                  4-hydroxyphenoxyacetic                                                                     7β-(4-hydroxyphenoxyacetamido)-                                          3-(1-methyltetrazol-5-ylthio-                                                 methyl)-Δ.sup.3 -0-2-isocephem-4-                                       carboxylic acid                                                  2,4-dichlorophenoxyacetic                                                                  7β-(2,4-dichlorophenoxy-                                    acid         acetamido)-3-(1-methyltetrazol-                                               5-ylthiomethyl)-Δ.sup.3 -0-2-isocephem-                                 4-carboxylic acid                                                2,6-dimethoxyphenoxyacetic                                                                 7β-(2,6-dimethoxyphenoxy-                                   acid         acetamido)-3-(1-methyltetrazol-                                               5-ylthiomethyl)-Δ.sup.3 -0-2-isocephem-                                 4-carboxylic acid                                                4-cyanophenoxyacetic acid                                                                  7β-(4-cyanophenoxyacetamido)-                                            3-(1-methyltetrazol-5-ylthio-                                                 methyl)-Δ.sup.3 -0-2-isocephem-4-                                       carboxylic acid                                                  α-phenoxypropionic acid                                                              7β-(α-phenoxypropionamido)-3-                                      (1-methyltetrazol-5-ylthiomethyl)-                                            Δ.sup.3 -0-2-isocephem-4-carboxylic                                     acid                                                             α-(2-chlorophenoxy)-                                                                 7β-[α-(2-chlorophenoxy)propion-                       propionic acid                                                                             amido]-3-(1-methyltetrazol-5-                                                 ylthiomethyl)-Δ.sup.3 -0-2-isocephem-                                   4-carboxylic acid                                                α-(2,4-dichlorophenoxy)-                                                             7β-[α-(2,4-dichlorophenoxy)-                          n-butyric acid                                                                             n-butyramido]-3-(1-methyltetrazol-                                            5-ylthiomethyl)-Δ.sup. 3 -0-2-isocephem-                                4-carboxylic acid                                                α-phenoxyphenylacetic                                                                7β-(α-phenoxyphenylacetamido)-                        acid         3-(1-methyltetrazol-5-ylthio-                                                 methyl)-Δ.sup.3 -0-2-isocephem-4-                                       carboxylic acid                                                  α-phenoxybutyric acid                                                                7β-(α-phenoxybutyramido)-3-                                        (1-methyltetrazol-5-ylthio-                                                   methyl)-Δ.sup.3 -0-2-isocephem-4-                                       carboxylic acid                                                  4-trifluoromethylphenoxy-                                                                  7β-(4-trifluoromethylphenoxy-                               acetic acid  acetamido)-3-(1-methyltetrazol-                                               5-ylthiomethyl)-Δ.sup.3 -0-2-                                           isocephem-4-carboxylic acid                                      benzyloxyacetyl chloride                                                                   7β-(benzyloxyacetamido)-3-                                               (1-methyltetrazol-5-ylthiomethyl)-                                            Δ.sup.3 -0-2-isocephem-4-carboxylic acid                   β-naphthoxyacetyl                                                                     7β-(β-naphthoxyacetamido)-3-                           chloride     (1-methyltetrazol-5-ylthiomethyl)-                                            Δ.sup.3 -0-2-isocephem-4-carboxylic acid                   __________________________________________________________________________

EXAMPLE 28

Following the general N-acylation methods of the preceeding examples,the compounds listed below are prepared by acylating7β-amino-3-(1-methyltetrazol-5-ylthiomethyl)-Δ³-0-2-isocephem-4-carboxylic acid with an acylating acid of the formula##STR143## or a functional equivalent, e.g. acid halide, thereof.

    ______________________________________                                         ##STR144##                                                                   R.sup.a          R.sup.b     R.sup.c                                          ______________________________________                                         ##STR145##                                                                    H                H                                                            H#STR146##       H                                                            H#STR147##       H                                                            H#STR148##       H                                                            H#STR149##       H                                                            H#STR150##       H                                                            H#STR151##       H                                                           4-pyridyl        H           H                                                3-pyridyl        H           H                                                 H#STR152##       H                                                            H#STR153##       H                                                           imidazolyl (2)   H           H                                                imidazolinyl (2) H           H                                                thiazolyl (2)    H           H                                                thiazolinyl (2)  H           H                                                triazolyl (2)    H           H                                                1-methyl-        H           H                                                 imidazolyl (2)                                                               2-thienyl        H           H                                                3-thienyl        H           H                                                n-butyl          H           H                                                isobutyl         H           H                                                2-acetamido-     H           H                                                 thiazol-5-yl                                                                 2-phenyl-1,3,4-  H           H                                                 thiadiazol-5-yl                                                              2-methyl-1,3,4-  H           H                                                 oxadiazol-5-yl                                                               ______________________________________                                    

EXAMPLE 29

Following the general N-acylation methods of the proceeding examples,the compounds listed below are prepared by acylation of7β-amino-3-(1-methyltetrazol5-ylthiomethyl)-Δ³-0-2-isocephem-4-carboxylic acid with the appropriate acylating acid ofthe general formula##STR154##7β-(3-phenyl-5-methyl-isoxazol-4-ylcarboxamido)-3-(1-methyltetrazol-5-ylthiomethyl)-Δ³-0-2-isocephem-4-carboxylic acid;

7β-[3-(2,6-dichlorophenyl)-5-methylisoxazol-4-ylcarboxamido]-3-(1-methyltetrazol-5-ylthiomethyl)-Δ³-0-2-isocephem-4-carboxylic acid;

7β-(2,6-dichlorobenzamido)-3-(1-methyltetrazol-5-ylthiomethyl)-Δ³-0-2-isocephem-4-carboxylic acid;

7β-(2-phenylbenzamido)-3-(1-methyltetrazol-5-ylthiomethyl)-Δ.sup.3-0-2-isocephem-4-carboxylic acid;

7β-(2-aminomethylbenzamido)-3-(1-methyltetrazol-5-ylthiomethyl)-.DELTA.³-0-2-isocephem-4-carboxylic acid;

7β-(2-carboxybenzamido)-3-(1-methyltetrazol -5-ylthiomethyl)-Δ³-0-2-isocephem-4-carboxylic acid;

7β-(cyclopentanecarboxamido)-3-(1-methyltetrazol-5-ylthiomethyl)-.DELTA.³-0-2-isocephem-4-carboxylic acid;

7β-(1-aminocyclohexanecarboxamido)-3-(1-methyltetrazol-5-ylthiomethyl)-Δ³-0-2-isocephem-4-carboxylic acid;

7β-(cyclohexanecarboxamido)-3-(1-methyltetrazol- 5-ylthiomethyl)-Δ³-0-2-isocephem-4-carboxylic acid;

7β-(1,4-cyclohexadienylcarboxamido)-3-(1-methyltetrazol-5-ylthiomethyl)-Δ³-0-2-isocephem-4-carboxylic acid;

7β-(4-nitrobenzamido)-3-(1-methyltetrazol-5-ylthiomethyl)-Δ.sup.3-0-2-isocephem-4-carboxylic acid;

7β-(4-methylbenzamido)-3-(1-methyltetrazol-5-ylthiomethyl)-Δ.sup.3-0-2-isocephem-4-carboxylic acid;

7β-(o-methoxybenzamido)-3-(1-methyltetrazol-5-ylthiomethyl)-Δ.sup.3-0-2-isocephem-4-carboxylic acid;

7β-(o-bromobenzamido)-3-(1-methyltetrazol-5-ylthiomethyl)-Δ.sup.3-0-2-isocephem-4-carboxylic acid;

7β-(p-ethoxybenzamido)-3-(1-methyltetrazol-5-ylthiomethyl)-Δ.sup.3-0-2-isocephem-4-carboxylic acid;

7β-(o-acetamidobenzamido)-3-(1-methyltetrazol-5-ylthiomethyl)-Δ.sup.3-0-2-isocephem-4-carboxylic acid;

7β-(p-allylbenzamido)-3-(1-methyltetrazol-5-ylthiomethyl)-Δ.sup.3-0-2-isocephem-4-carboxylic acid;

7β-(2,5-dihydroxybenzamido)-3-(1-methyltetrazol-5-ylthiomethyl)-.DELTA.³-0-2-isocephem-4-carboxylic acid;

7β-(2-ethoxy-1-naphthamido)-3-(1-methyltetrazol-5-ylthiomethyl)-.DELTA.³-0-2-isocephem-4-carboxylic acid;

7β-(2-methoxy-1-naphthamido)-3-(1-methyltetrazol-5-ylthiomethyl)-.DELTA.³-0-2-isocephem-4-carboxylic acid;

7β-(o-dimethylaminobenzamido)-3-(1-methyltetrazol-5-ylthiomethyl)-.DELTA.³-0-2-isocephem-4-carboxylic acid;

7β-(benzamido)-3-(1-methyltetrazol-5-ylthiomethyl)-Δ³-0-2-isocephem-4-carboxylic acid;

7β-(p-chlorobenzamido)-3-(1-methyltetrazol-5-ylthiomethyl)-Δ.sup.3-0-2-isocephem-4-carboxylic acid;

7β-(2-thienylcarboxamido)-3-(1-methyltetrazol-5-ylthiomethyl)-Δ.sup.3-0-2-isocephem- 4-carboxylic acid;

7β-(3-thienylcarboxamido)-3-(1-methyltetrazol-5-ylthiomethyl)-Δ.sup.3-0-2-isocephem-4-carboxylic acid;

7β-(2-furylcarboxamido)-3-(1-methyltetrazol-5-ylthiomethyl)-Δ.sup.3-0-2-isocephem-4-carboxylic acid;

7β-(3-furylcarboxamido)-3-(1-methyltetrazol-5-ylthiomethyl)-Δ.sup.3-0-2-isocephem-4-carboxylic acid;

7β-(2'-chlorocyclobutanecarboxamido)-3-(1-methyltetrazol-5-ylthiomethyl)-Δ³-0-2-isocephem-4-carboxylic acid;

7β-(3'-fluorocyclopentanecarboxamido)-3-(1-methyltetrazol-5-ylthiomethyl)-Δ³-0-2-isocephem-4-carboxylic acid;

7β-(3'-methylcyclopentanecarboxamido)-3-(1-methyltetrazol-5-ylthiomethyl)-Δ³-0-2-isocephem-4-carboxylic acid;

7β-(3'-methoxycyclopentanecarboxamido)-3-(1-methyltetrazol-5-ylthiomethyl)-Δ³-0-2-isocephem-4-carboxylic acid;

7β-(α-naphthamido)-3-(1-methyltetrazol-5-ylthiomethyl)-Δ.sup.3-0-2-isocephem-4-carboxylic acid;

7β-(β-naphthamido)-3-(1-methyltetrazol-5-ylthiomethyl)-Δ.sup.3-0-2-isocephem-4-carboxylic acid;

7β-(1-aminocyclopentanecarboxamido)-3-(1-methyltetrazol-5-ylthiomethyl)-Δ³-0-2-isocephem-4-carboxylic acid;

7β-(1-aminocycloheptanecarboxamido)-3-(1-methyltetrazol-5-ylthiomethyl)-Δ³-0-2-isocephem-4-carboxylic acid;

7β-(1-cyclohexenecarboxamido)-3-(1-methyltetrazol-5-ylthiomethyl)-.DELTA.³-0-2-isocephem-4-carboxylic acid.

EXAMPLE 30

Repeating the general N-acylation procedures of the above examples toreact trityl chloride with7β-amino-3-(1-methyltetrazol-5-ylthiomethyl)-Δ³-0-2-isocephem-4-carboxylic acid (or an ester or salt thereof), there isobtained after removal of any carboxyl-protecting group,7β-triphenylmethylcarboxamido-3-(1-methyltetrazol-5-ylthiomethyl)-.DELTA.³-0-2-isocephem-4-carboxylic acid.

EXAMPLE 31

Following the acylation methods of the preceeding examples and inparticular those disclosed in U.S. Pat. No. 3,546,219, the compoundslisted below are prepared by reacting7β-amino-3-(1-methyltetrazol-5-ylthiomethyl)-Δ³-0-2-isocephem-4-carboxylic acid (or an ester or salt thereof), with theappropriate acylating agent. ##STR155##

EXAMPLE 32

Following the acylation methods of the preceeding examples and inparticular those disclosed in U.K. Patents 1,296,081 and 1,294,541, thecompounds listed below are prepared by reacting7β-amino-3-(1-methyltetrazol-5-ylthiomethyl)-Δ³-0-2-isocephem-4-carboxylic acid or an ester or salt thereof with anacylating agent of the formula ##STR156## or a functional equivalentthereof. ##STR157##

EXAMPLE 33

When 7β-amino-3-(1-methyltetrazol-5-ylthiomethyl)-Δ³-0-2-isocephem-4-carboxylic acid or an ester or salt thereof is acylatedaccording to the procedures above and in particular those disclosed inU.S. Pat. No. 3,692,779 with an acid chloride of the formula ##STR158##there are produced the compounds listed below. ##STR159##dichloromethyl; n-propyl;

cyclopentyl;

cyclohexyl;

p-chlorobenzyl;

phenyl;

2-thienyl;

3-thienyl.

EXAMPLE 34

When the 7-acylamido-3-(1-methyltetrazol-5-ylthiomethyl)-Δ³-0-2-isocephem-4-carboxylic acid compounds of Example 33 arehydrogenated as by the process of U.S. Pat. No. 3,692,779, there areproduced the compounds listed below. ##STR160## where R^(i) is asdefined in Example 33.

EXAMPLE 35

When 7β-amino-3-(1-methyltetrazol-5-ylthiomethyl)-Δ³-0-2-isocephem-4-carboxylic acid or an ester or salt thereof is acylatedaccording to the procedures of the above examples (and in particular theprocedures disclosed in U.S. Pat. No. 3,646,024) with an acid chlorideof the formula ##STR161## there are produced the compounds listed below.##STR162##

EXAMPLE 36

When 7β-amino-3-(1-methyltetrazol-5-ylthiomethyl)-Δ³-0-2-isocephem-4-carboxylic acid or an ester or salt thereof is acylatedaccording to the procedures above and in particular according to themethods of U.S. Pat. No. 3,778,436 with an acylating agent of theformula ##STR163## or a functional equivalent thereof, there areproduced the compounds listed below

    ______________________________________                                         ##STR164##                                                                          R.sup.a          R.sup.1                                               ______________________________________                                        3,4-dimethoxyphenyl    ethyl                                                  p-methylphenyl         2-thienyl                                              2,4-dichlorophenyl     ethyl                                                  5-methyl-3-phenyl-     ethyl                                                   isoxazol-4-yl                                                                2-thienyl              ethyl                                                  2-furyl                2-furyl                                                phenyl                 phenyl                                                 1,4-cyclohexadien-     methyl                                                  1-yl                                                                         ______________________________________                                    

EXAMPLE 37

When p-nitrobenzyl 7β-amino-3-(1-methyltetrazol-5-ylthiomethyl)-Δ³-0-2-isocephem-4-carboxylate is reacted with equimolar quantities ofEEDQ and the N-t-butoxycarbonylamino acylating acids listed below, thereare produced after catalytic hydrogenation the following compounds.

    ______________________________________                                        Acylating Agent   Product                                                     ______________________________________                                        D-(-)-α-(3,5-dichloro-4-                                                                7β-[D-(-)-α-amino-α-(3,5-                    hydroxyphenyl)-α-(t-butoxy-                                                             dichloro-4-hydroxyphenyl)-                                    carbonylamino)acetic acid                                                                     acetamido]-3-(1-methyl-                                                       tetrazol-5-ylthiomethyl)-Δ.sup.3 -                                      0-2-isocephem-4-carboxylic acid                               D-(-)-α-(3-chloro-4-                                                                    7β-[D-(-)-α-amino-α-(3-chloro-               hydroxyphenyl)-2-(t-butoxy-                                                                   4-hydroxyphenyl)acetamido]-3-                                 carbonylamino)acetic acid                                                                     (1-methyltetrazol-5-ylthiomethyl)-                                            Δ.sup.3 -0-2-isocephem-4-carboxylic                                     acid                                                          D-(-)-α-(p-hydroxyphenyl)-                                                              7β-[D-(-)-α-amino-α-(p-hydroxy-              α-(t-butoxycarbonylamino)-                                                              phenyl)acetamido]-3-(1-methyl-                                acetic acid     tetrazol-5-ylthiomethyl)-Δ.sup.3 -                                      0-2-isocephem-4-carboxylic acid                               2-(N-t-butoxycarbonyl-                                                                        7β-[α-aminoα-(2,4,6-cyclo-                   amino)-2-(2,4,6-cyclo-                                                                        heptatrien-1-yl)acetamido]-3-                                 heptatrien-1-yl acetic                                                                        (1-methyltetrazol-5-ylthiomethyl)-                            acid            Δ.sup.3 -0-2-isocephem-4-carboxylic acid                D-2-(t-butoxycarbonyl-                                                                        7β-[D-α-amino-α-(3'-hydroxy-                 amino)-2-(3'-hydroxy-                                                                         phenyl)acetamido]-3-(1-methyl-                                phenyl)acetic acid                                                                            tetrazol-5-ylthiomethyl)-Δ.sup.3 -0-2-                                  isocephem-4-carboxylic acid                                   D-α-(t-butoxycarbonyl-                                                                  7β-[D-α-amino-α-(4-acetamido-                amino)-α-4-acetamido-                                                                   phenyl)acetamido]-3-(1-methyl-                                phenylacetic acid                                                                             tetrazol-5-ylthiomethyl)-Δ.sup.3 -                                      0-2-isocephem-4-carboxylic acid                               D-2-(t-butoxycarbonyl-                                                                        7β-[D-α-amino-α-(1,4-cyclo-                  amino)-2-(1,4-cyclo-                                                                          hexadienyl)acetamido]-3-(1-                                   hexadienyl)acetic acid                                                                        methyltetrazol-5-ylthiomethyl)-                                               Δ.sup.3 -0-2-isocephem-4-carboxylic                                     acid                                                          D-3-(t-butoxycarbonyl-                                                                        7β-[D-3'-amino-3'-(1,4-cyclo-                            amino)-3-(1,4-cyclo-                                                                          hexadienyl)propionamido]-3-(1-                                hexadienyl)propionic acid                                                                     methyltetrazol-5-ylthiomethyl)-                                               Δ.sup.3 -0-2-isocephem-4-carboxylic                                     acid                                                          D-(-)-α-(2-thienyl)-α  -                                                          7β-[D-α-amino-α-(2-thienyl)-                 (t-butoxycarbonylamino)-                                                                      acetamido]-3-(1-methyltetrazol-                               acetic acid     5-ylthiomethyl)-Δ.sup.3 -0-2-isocephem-                                 4-carboxylic acid                                             D-(-)-α-(3-thienyl)-α-                                                            7β-[D-α-amino-α-(3-thienyl)-                 (t-butoxycarbonylamino)-                                                                      acetamido]-3-(1-methyltetrazol-                               acetic acid     5-ylthiomethyl)-Δ.sup.3 -0-2-isocephem-                                 4-carboxylic acid                                             D-α-(t-butoxycarbonyl-                                                                  7β-[D-α-amino-α-(o-hydroxy-                  amino)-α-(o-hydroxyphenyl)-                                                             phenyl)acetamido]-3-(1-methyl-                                acetic acid     tetrazol-5-ylthiomethyl)-Δ.sup.3 -                                      0-2-isocephem-4-carboxylic acid                               D-α-(t-butoxycarbonyl-                                                                  7β-[D-α-amino-α-(p-nitro-                    amino)-α-(p-nitrophenyl)-                                                               phenyl)acetamido]-3-(1-methyl-                                acetic acid     tetrazol-5-ylthiomethyl)-Δ.sup.3 -                                      O-2-isocephem-4-carboxylic 0-2-isocephem-4-carboxylic                         acid                                                          D-α-(t-butoxycarbonyl-                                                                  7β-[D-α-amino-α-(p-methoxy-                  amino)-α-(p-methoxyphenyl)-                                                             phenyl)acetamido]-3-(1-methyl-                                acetic acid     tetrazol-5-ylthiomethyl)-Δ.sup.3 -                                      0-2-isocephem-4-carboxylic acid                               D-α-(t-butoxycarbonyl-                                                                  7β-[D-α-amino-α-(p-cyanophenyl)-             amino)-α-(p-cyanophenyl)-                                                               acetamido]-3-(1-methyltetrazol-                               acetic acid     5-ylthiomethyl)-Δ.sup.3 -0-2-isocephem-                                 4-carboxylic acid                                             D-α-(t-butoxycarbonyl-                                                                  7β-[D-α-amino-α-(p-methylthio-               amino)-α-(p-methylthio-                                                                 phenyl)acetamido]-3-(1-methyltetra-                           phenyl)acetic acid                                                                            zol-5-ylthiomethyl)-Δ.sup.3 -0-2-                                       isocephem-4-carboxylic acid                                   D-α-(t-butoxycarbonyl-                                                                  7β-[D-α-amino-α-(p-isopropyl-                amino-α-(p-isopropyl-                                                                   phenyl)acetamido]-3-(1-methyl-                                phenyl)acetic acid                                                                            tetrazol-5-ylthiomethyl)-Δ.sup.3 -0-2-                                  isocephem-4-carboxylic acid                                   D-α-(t-butoxycarbonyp-                                                                  7β-[D-α-amino-α-(o-sulfamyl-                 amino)-α-(o-sulfamyl-                                                                   phenyl)acetamido]-3-(1-methyl-                                phenyl)acetic acid                                                                            tetrazol-5-ylthiomethyl)-Δ.sup.3 -                                      0-2-isocephem-4-carboxylic acid                               D-α-(t-butoxycarbonyl-                                                                  7β-[D-α-aino-α-(o-amino-                     amino)-α-(o-aminomethyl-                                                                methylphenyl)acetamido]-3-(1-                                 phenyl)acetic acid                                                                            methyltetrazol-5-ylthiomethyl)-                                               Δ.sup.3 -0-2-isocephem-4-carboxylic                                     acid                                                          D-α-(t-butoxycarbonyl-                                                                  7β-[D-α-amino-α-(o-dimethyl-                 amino)-α-(o-dimethylamino-                                                              aminophenyl)acetamido]-3-(1-                                  phenyl)acetic acid                                                                            methyltetrazol-5-ylthiomethyl)-                                               Δ.sup.3 -0-2-isocephem-4-carboxylic                                     acid                                                          D-α-(t-butoxycarbonyl-                                                                  7β-[D-α-amino-α-(4-chloro-2-                 amino)-α-(4-chloro-2-                                                                   thienyl)acetamido]-3-(1-methyl-                               thienyl)acetic acid                                                                           tetrazol-5-ylthiomethyl)-Δ.sup.3 -                                      0-2-isocephem-4-carboxylic acid                               D-α-(t-butoxycarbonyl-                                                                  7β-[D-α-amino-α-(cyclohexyl)-                amino)-α-(cyclohexyl)-                                                                  acetamido]-3-(1-methyltetrazol-                               acetic acid     5-ylthiomethyl)-Δ.sup.3 -0-2-isocephem-                                 4-carboxylic acid                                             D-α-(t-butoxycarbonyl-                                                                  7β-[D-α-amino-α-(3-trifluoro-                amino)-α-(3-trifluoro-                                                                  methylphenyl)acetamido]-3-(1-                                 methylphenyl)acetic acid                                                                      methyltetrazol-5-ylthiomethyl)-                                               Δ.sup.3 -0-2-isocephem-4-carboxylic                                     acid                                                          ______________________________________                                    

EXAMPLE 38

When 7β-amino-3-(1-methyltetrazol-5-ylthiomethyl)-Δ³-0-2-isocephem-4-carboxylic acid or an ester or salt thereof is acylatedaccording to the general procedures of the preceeding examples with theacylating agents listed below (suitably protected), there are producedthe following compounds.

    ______________________________________                                        Acylating Agent   Product                                                     ______________________________________                                        α-amino-α-(1-cyclohexenyl)-                                                       7β-[α-amino-α-(1-cyclohexenyl)-              acetic acid     acetamido]-3-(1-methyltetrazol-                                               5-ylthiomethyl)-Δ.sup.3 -0-2-isocephem-                                 4-carboxylic acid                                             α-amino-α-(isothiazol-4-                                                          7β-[α-amino-α-(isothiazol-4- -yl)acetyl                      chloride yl)acetamido]-3-(1-methyltetrazol-                                   5-ylthiomethyl)-Δ.sup.3 -0-2-isocephem-                                 4-carboxylic acid                                             α-amino-α-(5-phenyl-1,3,4-                                                        7β-[α-amino-α-(5-phenyl-1,3,4-               thiadiazol-2-yl)acetyl                                                                        thiadiazol-2-yl)acetamido]-3-                                 chloride        (1-methyltetrazol-5-ylthiomethyl)-                                            Δ.sup.3 -0-2-isocephem-4-carboxylic acid                α-amino-α-(5-phenyl-1,3,4-                                                        7β-[α-amino-α-(5-phenyl-1,3,4-               oxadiazol-2-yl)acetyl                                                                         oxadiazol-2-yl)acetamido]-3-                                  chloride        (1-methyltetrazol-5-ylthiomethyl)-                                            Δ.sup.3 -0-2-isocephem-4-carboxylic acid                α-amino-α-(3-methyl-1,2,5-                                                        7β[α-amino-α-(3-methyl-1,2,5-                oxadiazol-4-yl)acetyl                                                                         oxadiazol-4-yl)acetamido]-3-                                  chloride        (1-methyltetrazol-5-ylthiomethyl)-                                            Δ.sup.3 -0-2-isocephem-4-carboxylic acid                α-amino-α-(oxazol-2-yl)-                                                          7β-[α-amino-α-(oxazol-2-yl)-                 acetyl chloride acetamido]-3-(1-methyltetrazol-                                               5-ylthiomethyl)-Δ.sup.3 -0-2-isocephem-                                 4-carboxylic acid                                             α-amino-α-(1H)-tetra-                                                             7β-[α-amino-α-(1H)-tetrazolyl-               zolylacetyl chloride                                                                          acetamido]-3-(1-methyltetrazol-                                               5-ylthiomethyl)-Δ.sup.3 -0-2-isocephem-                                 4-carboxylic acid                                             α-amino-α-4-isoxazolyl-                                                           7β-[α-amino-α-(4-isoxazolyl)-                acetyl chloride acetamido]-3-(1-methyltetrazol-                                               5-ylthiomethyl)-Δ.sup.3 -0-2-isocephem-                                 4-carboxylic acid                                             α-amino-α-(2-thiazolyl)-                                                          7β-[α-amino-α-(2-thiazolyl)-                 acetyl chloride acetamido]-3-(1-methyltetrazol-                                               5-ylthiomethyl)-Δ.sup.3 -0-2-isocephem-                                 4-carboxylic acid                                             α-amino-α-(2-furyl)-                                                              7β-[α -amino-α-(2-furyl)-                    acetyl chloride acetamido]-3-(1-methyltetrazol-                                               5-ylthiomethyl)-Δ.sup.3 -0-2-isocephem-                                 4-carboxylic acid                                             α-amino-α-(1,2,5-thiadiazol-                                                      7β-[α-amino-α-(1,2,5-thiadizaol-             3-yl)acetyl chloride                                                                          3-yl)acetamido]-3-(1-methyl-                                                  tetrazol-5-ylthiomethyl)-Δ.sup.3 -0-2-                                  isocephem-4-carboxylic acid                                   α-amino-α-(3-furyl)acetyl                                                         7β-[α-amino-α-(3-furyl)-                     chloride        acetamido]-3-(1-methyltetrazol-                                               5-ylthiomethyl)-Δ.sup.3 -0-2-isocephem-                                 4-carboxylic acid                                             ______________________________________                                    

EXAMPLE 39

When benzyl 7β-amino-3-(1-methyltetrazol-5-ylthiomethyl)-Δ³-0-2-isocephem-4-carboxylate is acylated according to the generalacylation procedures above with the acylating agents listed below(suitably protected if necessary), there are produced the followingcompounds after removal of any protecting groups:

    ______________________________________                                        Acylating Agent   Product                                                     ______________________________________                                        D-3-chloromandelic acid                                                                       7β-[D-α-hydroxy-α-(3-chloro-                                 phenyl)acetamido]-3-(1-methyl-                                                tetrazol-5-ylthiomethyl)-Δ.sup.3 -                                      0-2-isocephem-4-carboxylic acid                               D-2-trifluoromethyl-                                                                          7β-[D-α-hydroxy-α-(2-trifluoro-              mandelic acid   methylphenyl)acetamido]-3-(1-                                                 methyltetrazol-5-ylthiomethyl)-                                               Δ.sup.3 -0-2-isocephem-4-carboxylic                                     acid                                                          D-3-nitromandelic acid                                                                        7β-[D-α-hydroxy-α-(3-nitrophenyl)                            acetamido -3-(1-methyltetrazol-                                               5-ylthiomethyl)-Δ.sup.3 -0-2-isocephem-                                 4-carboxylic acid                                             D-p-hydroxymandelic acid                                                                      7β-[D-α-hydroxy-α-(p-hydroxy-                                phenyl)acetamido]-3-(1-methyl-                                                tetrazol-5-ylthiomethyl)-Δ.sup.3 -                                      0-2-isocephem-4-carboxylic acid                               D-3-chloro-4-hydroxy-                                                                         7β-[D-α-hydroxy-α-(3-chloro-4-               mandelic acid   hydroxyphenyl)acetamido]-3-(1-                                                methyltetrazol-5-ylthiomethyl)-                                               Δ.sup.3 -0-2-isocephem-4-carboxylic                                     acid                                                          D-3,5-dichloro-4-hydroxy-                                                                     7β-[D-α-hydroxy-α-(3,5-dichloro-             mandelic acid   4-hydroxyphenyl)acetamido]-3-                                                 (1-methyltetrazol-5-ylthiomethyl)-                                            Δ.sup.3 -0-2-isocephem-4-carboxylic acid                D-o-methylaminomandelic                                                                       7β-[D-α-hydroxy-α-(o-methyl-                 acid            aminophenyl)acetamido]-3-(1-                                                  methyltetrazol-5-ylthiomethyl)-                                               Δ.sup.3 -0-2-isocephem-4-carboxylic acid                D-p-methoxymandelic acid                                                                      7β-[D-α-hydroxy-α-(p-methoxy-                                phenyl)acetamido]-3-(1-methyl-                                                tetrazol-5-ylthiomethyl)-Δ.sup.3 -0-2-                                  isocephem-4-carboxylic acid                                   D-m-methylthiomandelic                                                                        7β-[D-α-hydroxy-α-(m-methyl-                 acid            thiophenyl)acetamido]-3-(1-                                                   methyltetrazol-5-ylthiomethyl)-                                               Δ.sup.3 -0-2-isocephem-4-carboxylic                                     acid                                                          D-m-iodomandelic acid                                                                         7β-[D-α-hydroxy-α-(m-iodophenyl)                             acetamido]-3-(1-methyltetrazol-                                               5-ylthiomethyl)-Δ.sup.3 -0-2-isocephem-                                 4-carboxylic acid                                             4-isoxazoleglycolic acid                                                                      7β-[α-hydroxy-α-(4-isoxazolyl)-                              acetamido]-3-(1-methyltetrazol-                                               5-ylthiomethyl)-Δ.sup.3 -0-2-isocephem-                                 4-carboxylic acid                                             4-thiazoleglycolic acid                                                                       7β-[α-hydroxy-α-(4-thiazolyl)-                               acetamido]-3-(1-methyltetrazol-                                               5-ylthiomethyl)-Δ.sup.3 -0-2-isocephem-                                 4-carboxylic acid                                             4-oxazoleglycolic acid                                                                        7β-[α-hydroxy-α-(4-oxazolyl)-                                acetamido]-3-(1-methyltetrazol-                                               5-ylthiomethyl)-Δ.sup.3 -0-2-isocephem-                                 4-carboxylic acid                                             3-isothiazoleglycolic acid                                                                    7β-[α-hydroxy-α-(3-isothiazolyl)-                            acetamido]-3-(1-methyltetrazol-                                               5-ylthiomethyl)-Δ.sup.3 -0-2-isocephem-                                 4-carboxylic acid                                             1,2,3-triazole-4-glycolic                                                                     7β-[α-hydroxy-α-(1,2,3-triazol-              acid            4-yl)acetamido]-3-(1-methyl-                                                  tetrazol-5-ylthiomethyl)-Δ.sup.3 -0-2-                                  isocephem-4-carboxylic acid                                   5-isoxazoleglycolic acid                                                                      7β-[α-hydroxy-α-(5-isoxazolyl)-                              acetamido]-3-(1-methyltetrazol-                                               5-ylthiomethyl)-Δ.sup.3 -0-2-isocephem-                                 4-carboxylic acid                                             1,2,4-triazole-3-glycolic                                                                     7β-[α-hydroxy-α-(1,2,4-triazol-              acid            3-yl)acetamido]-3-(1-methyltetra-                                             zol-5-ylthiomethyl)-Δ.sup.3 -0-2-                                       isocephem-4-carboxylic acid                                   2-thienylglycolic acid                                                                        7β-[α-hydroxy-α-(2-thienyl)-                                 acetamido]-3-(1-methyltetrazol-                                               5-ylthiomethyl)-Δ.sup.3 -0-2-isocephem-                                 4-carboxylic acid                                             3-thienylglycolic acid                                                                        7β-[α-hydroxy-α-(3-thienyl)-                                 acetamido]-3-(1-methyltetrazol-                                               5-ylthiomethyl)-Δ.sup.3 -0-2-isocephem-                                 4-carboxylic acid                                             1,4-cyclohexadien-1-yl-                                                                       7β-[α-hydroxy-α-(1,4-cyclo-                  glycolic acid   hexadien-1-yl)acetamido]-3-                                                   (1-methyltetrazol-5-ylthiomethyl)-                                            Δ.sup.3 -0-2-isocephem-4-carboxylic acid                1-cyclohexenylglycolic                                                                        7β-[α-hydroxy-α-(1-cyclo-                    acid            hexenyl)acetamido]-3-(1-methyl-                                               tetrazol-5-ylthiomethyl)-Δ.sup.3 -                                      0-2-isocephem-4-carboxylic acid                               2-pyrrolylglycolic acid                                                                       7β-[α-hydroxy-α-(2-pyrrolyl)-                                acetamido]-3-(1-methyltetrazol-                                               5-ylthiomethyl)-Δ.sup.3 -0-2-isocephem-                                 4-carboxylic acid                                             2-furylglycolic acid                                                                          7β-[α-hydroxy-α-(2-furyl)-                                   acetamido]-3-(1-methyltetrazol-                                               5-ylthiomethyl)-Δ.sup.3 -0-2-isocephem-                                 4-carboxylic acid                                             3-pyridylglycolic acid                                                                        7β-[α-hydroxy-α-(3-pyridyl)-                                 acetamido]-3-(1-methyltetrazol-                                               5-ylthiomethyl)-Δ.sup.3 -0-2-isocephem-                                 4-carboxylic acid                                             ______________________________________                                    

EXAMPLE 40

When benzyl 7β-amino-3-(1-methyltetrazol-5-ylthiomethyl)-Δ³-0-2-isocephem-4-carboxylate is acylated according to the generalacylation procedures described above with the acylating agents listedbelow (suitably protected if desired), there are produced the followingcompounds after removal of any protecting groups:

    ______________________________________                                        Acylating Agent  Product                                                      ______________________________________                                        p-hydroxyphenylmalonic                                                                       7β-[α-carboxy-α-(p-hydroxy-                   acid           phenyl)acetamido]-3-(1-methyl-                                                tetrazol-5-ylthiomethyl)-Δ.sup.3 -                                      0-2-isocephem-4-carboxylic acid                                3-chloro-4-hydroxyphenyl-                                                                    7β-[α-carboxy-α-(3-chloro-4-                  malonic acid   hydroxyphenyl)acetamido]-3-                                                   (1-methyltetrazol-5-ylthiomethyl)-                                            Δ.sup.3 -0-2-isocephem-4-carboxylic acid                 3,5-dichloro-4-hydroxy-                                                                      7β-[α-carboxy-α-(3,5-dichloro-                phenylmalonic acid                                                                           4-hydroxyphenyl)acetamido]-3-                                                 (1-methyltetrazol-5-ylthiomethyl)-                                            Δ.sup.3 -0-2-isocephem-4-carboxylic acid                 o-chlorophenylmalonic                                                                        7β-[α-carboxy-α-(o-chloro-                    acid           phenyl)acetamido]-3-(1-methyl-                                                tetrazol-5-ylthiomethyl)-Δ.sup.3 -                                      0-2-isocephem-4-carboxylic acid                                p-nitrophenylmalonic                                                                         7β-[α-carboxy-α-(p-nitrophenyl)               acid           acetamido]-3-(1-methyltetrazol-                                               5-ylthiomethyl)-Δ.sup.3 -0-2-isocephem-                                 4-carboxylic acid                                              p-acetoxyphenylmalonic                                                                       7β-[α-carboxy-α-(p-acetoxyphenyl)             acid           acetamido]-3-(1-methyltetrazol-                                               5-ylthiomethyl)-Δ.sup.3 -0-2-isocephem-                                 4-carboxylic acid                                              p-methoxyphenylmalonic                                                                       7β-[α-carboxy-α-(p-methoxy-                   acid           phenyl)acetamido]-3-(1-methyl-                                                tetrazol-5-ylthiomethyl)-Δ.sup.3 -                                      0-2-isocephem-4-carboxylic acid                                p-methylthiophenylmalonic                                                                    7β-[α-carboxy-α-(p-methylthiophenyl)          acid           acetamido]-3-(1-methyltetrazol-                                               5-ylthiomethyl)-Δ.sup.3 -0-2-isocephem-                                 4-carboxylic acid                                              p-cyanophenylmalonic                                                                         7β-[α-carboxy-α-(p-cyanophenyl)-              acid           acetamido]-3-(1-methyltetrazol-                                               5-ylthiomethyl)-Δ.sup.3 -0-2-isocephem-                                 4-carboxylic acid                                              m-isopropylphenylmalonic                                                                     7β-[α-carboxy-α-(m-isopropyl-                 acid           phenyl)acetamido]-3-(1-methyl-                                                tetrazol-5-ylthiomethyl)-Δ.sup.3 -                                      0-2-isocephem-4-carboxylic acid                                o-aminomethylphenyl-                                                                         7β-[α -carboxy-α-(o-aminomethyl-              malonic acid   phenyl)acetamido]-3-(1-methyl-                                                tetrazol-5-ylthiomethyl)-Δ.sup.3 -                                      0-2-isocephem-4-carboxylic acid                                o-dimethylaminophenyl-                                                                       7β-[α-carboxy-α-(o-dimethyl-                  malonic acid   aminophenyl)acetamido]-3-(1-                                                  methyltetrazol-5-ylthiomethyl)-                                               Δ.sup.3 -0-2-isocephem-4-carboxylic                                     acid                                                           2-thienylmalonic acid                                                                        7β-[60 -carboxy-α-(2-theinyl)-                                     acetamido]-3-(1-methyltetrazol-                                               5-ylthiomethyl)-Δ.sup.3 -0-2-isocephem-                                 4-carboxylic acid                                              3-thienylmalonic acid                                                                        7β-[α-carboxy-α-(3-thienyl)-                                 acetamido]-3-(1-methyltetrazol-                                               5-ylthiomethyl)-Δ.sup.3 -0-2-isocephem-                                 4-carboxylic acid                                              1,4-cyclohexadienyl-                                                                         7β-[α-carboxy-α-(1,4-cyclo-                   malonic acid   hexadien-1-yl)acetamido]-3-                                                   (1-methyltetrazol-5-ylthiomethyl)-                                            Δ.sup.3 -0-2-isocephem-4-carboxylic acid                 1-cyclohexenylmalonic                                                                        7β-[α-carboxy-α-(1-cyclohexenyl)              acid           acetamido]-3-(1-methyltetrazol-                                               5-ylthiomethyl)-Δ.sup.3 -0-2-isocephem-                                 4-carboxylic acid                                              2-furylmalonic acid                                                                          7β-[α-carboxy-α-(2-furyl)-                                   acetamido]-3-(1-methyltetrazol-                                               5-ylthiomethyl)-Δ.sup.3 -0-2-isocephem-                                 4-carboxylic acid                                              4-pyridylmalonic acid                                                                        7β-[α-carboxy-α-(4-pyridyl)-                                 acetamido]-3-(1-methyltetrazol-                                               5-ylthiomethyl)-Δ.sup.3 -0-2-isocephem-                                 4-carboxylic acid                                              ______________________________________                                    

EXAMPLE 41

When 7β-amino-3-(1-methyltetrazol-5-ylthiomethyl)-Δ³-0-2-isocephem-4-carboxylic acid or an ester or salt thereof is acylatedaccording to the general procedures of the preceeding examples with anacylating agent of the formula ##STR165## or a functional equivalentthereof, there are produced the compounds listed below

    ______________________________________                                         ##STR166##                                                                   R.sup.a             Y                                                         ______________________________________                                        phenyl             guanidino                                                  2-thienyl          guanidino                                                  3-thienyl          guanidino                                                  1,4-cyclohexadienyl                                                                              guanidino                                                  1-cyclohexenyl     guanidino                                                  p-hydroxyphenyl    guanidino                                                  3-chloro-4-hydroxyphenyl                                                                         guanidino                                                  3,5-dichloro-4-hydroxyphenyl                                                                     guanidino                                                  phenyl             ureido                                                     2-thienyl          ureido                                                     3-thienyl          ureido                                                     1-cyclohexenyl     ureido                                                     1,4-cyclohexadienyl                                                                              ureido                                                     p-hydroxyphenyl    ureido                                                     3,5-dichloro-4-hydroxyphenyl                                                                     ureido                                                     o-aminomethylphenyl                                                                              ureido                                                     p-methylphenyl     ureido                                                     m-chlorophenyl     ureido                                                     phenyl             thioureido                                                 2-thienyl          thioureido                                                 3-thienyl          thioureido                                                 1-cyclohexenyl     thioureido                                                 1,4-cyclohexadienyl                                                                              thioureido                                                 p-hydroxyphenyl    thioureido                                                 3-chloro-4-hydroxyphenyl                                                                         thioureido                                                 3,5-dichloro-4-hydroxyphenyl                                                                     thioureido                                                 o-aminomethylphenyl                                                                              methylthioureido                                           m-chlorophenyl     allylthioureido                                            phenyl             chloro                                                     2-thienyl          bromo                                                      3-thienyl          chloro                                                     1,4-cyclohexadienyl                                                                              methoxy                                                    1-cyclohexenyl     ethoxy                                                     phenyl             phenyl                                                     2-thienyl          methoxy                                                    3-thienyl          ethoxy                                                     p-hydroxyphenyl    iodo                                                       p-trifluoromethylphenyl                                                                          methoxy                                                    3,4-dichlorophenyl methoxy                                                    phenyl             acetoxy                                                    2-thienyl          acetoxy                                                    2-furyl            acetoxy                                                    p-nitrophenyl      acetoxy                                                    p-methoxyphenyl    acetoxy                                                    phenyl             propionyloxy                                               phenyl             cyano                                                      phenyl             SO.sub.3 H                                                 phenyl             azido                                                      phenyl             methylsulfonyl                                             phenyl             5-indanyloxycarbonyl                                       p-hydroxyphenyl    5-indanyloxycarbonyl                                       3-chloro-4-hydroxyphenyl                                                                         5-indanyloxycarbonyl                                       3,5-dichloro-4-hydroxyphenyl                                                                     5-indanyloxycarbonyl                                       ______________________________________                                    

EXAMPLE 42

Preparation of7β-[α-(2-Aminomethyl-1,4-cyclohexadienyl)acetamido]-3-(1-methyltetrazol-5-ylthiomethyl)-Δ³-0-2-isocephem-4-carboxylic acid ##STR167## A.α-(2-Aminomethyl-1,4-cyclohexadienyl)acetic acid

A solution of 16.5 g. (0.1 mole) of o-aminomethylphenylacetic acid in1.5 l of liquid ammonia (which had been treated with 50 mg. of Li toremove a trace of moisture) was slowly diluted with 500 ml. of dryt-BuOH. To the solution was added in small portions 3.4 g. (0.5 g.atom)of Li over a period of 4 hours and the mixture was stirred for 16 hoursat room temperature removing the liquid ammonia in a hood and finallyevaporated to dryness below 40° C. The residue was dissolved in 500 ml.of water and the solution was chromatographed on a column of IR-120 (H⁺,700 ml.) resin and eluted with 1% NH₄ OH solution. Ninhydrin positivefractions of the eluate were combined and evaporated to dryness. Theresidue was washed with four 50 ml. portions of hot acetone andrecrystallized from 500 ml. of ethanol-water (1:1) to give 11.2 g. (67%)of α-(2-aminomethyl-1,4-cyclohexadienyl)acetic acid as colorlessneedles. M.p. 183° C.

Ir: ν _(max) ^(nuj) 1630, 1520, 1380, 1356 cm⁻ ¹. ##STR168## Anal.Calc'd. cd. for C₉ H₁₃ NO₂ : C, 64.65; H, 7.84; N, 8.38. Found: C,64.77; H, 8.06; N, 8.44.

B. α-[2-(t-Butoxycarbonylaminomethyl)-1,4-cyclohexadienyl]-acetic acid

To a stirred solution of 8.0 g. (0.048 mole) ofα-(2-aminomethyl-1,4-cyclohexadienyl)acetic acid and 3.8 g. (.096 mole)of NaOH in 150 ml. of water was added a solution of 10.3 g. (0.072 mole)of t-butoxycarbonylazide in 80 ml. of THF and the mixture was stirredfor 18 hours at room temperature. The THF was removed under reducedpressure and the residual solution was washed with ether (2 × 100 ml.),acidified with 6 N HCl and extracted with ether (3 × 100 ml.). Thecombined extracts were washed with water (2 × 100 ml.) and a saturatedNaCl solution (100 ml.), dried with Na₂ SO₄ and evaporated to dryness.The oily residue was triturated with n-hexane to give 10.5 g. (82%) ofcolorless powder melting at 113° C.

Ir: ν _(max) ^(nuj) 3370, 1715, 1640, 1530, 1280, 1160 cm.sup.⁻¹.##STR169## Anal. Calc'd. for C₁₄ H₂₁ NO₄ : C, 62.90; H, 7.92; N, 5.24.Found: C, 63.13; H, 8.21; N, 5.26.

C.7β-[α-t-Butoxycarbonylaminomethyl-1,4-cyclohexadienyl)-acetamido]-3-(1-methyltetrazol-5-ylthiomethyl)-Δ³-0 -2-isocephem-4-carboxylic acid

To a stirred solution of equimolar amounts ofα-[2-(t-butoxycarbonylaminomethyl)-1,4-cyclohexadienyl)acetic acid and2,4-dinitrophenol in ethyl acetate is added an equimolar amount ofN,N'-dicyclohexylcarbodiimide. The reaction mixture is stirred at roomtemperature for 3 hours. The separated dicyclohexylurea is filtered off.The filtrate is evaporated to dryness to give the activated ester whichis dissolved in tetrahydrofuran. To this solution is added a solution of7β-amino-3-(1-methyltetrazol-5-ylthiomethyl)-Δ³-0-2-isocephem-4-carboxylic acid and triethylamine in approximately a1:2 molar proportion, respectively, relative to theα-[2-(t-butoxycarbonylaminomethyl)-1,4-cyclohexadienyl]acetic acid. Themixture is stirred at room temperature for several hours andconcentrated in vacuo. The concentrate is washed with ether, acidifiedwith dilute mineral acid and estracted with ethyl acetate. The extractsare washed with water and saturated NaCl solution and dried to give thetitle product.

D.7β-[α-(2-aminomethyl-1,4-cyclohexadienyl)acetamido]-3-(1-methyltetrazol-5-ylthiomethyl)-Δ³-0-2-isocephem-4-carboxylic acid

A solution of7β-[α-(2-t-butoxycarbonylaminomethyl-1,4-cyclohexadienyl)acetamido]-3-(1-methyltetrazol-5-ylthiomethyl)-Δ³-0-2-isocephem-4-carboxylic acid in trifluoroacetic acid is stirred at0° C. for 1 hour. To the solution is added dry ether until a precipitateforms. The precipitate is collected by filtration, suspended in waterand adjusted to pH6 to give the title product.

Example 43

7β-[α-(2-Aminomethyl-1-cyclohexenyl)acetamido]-3-(1-methyltetrazol-5-ylthiomethyl)-Δ³-0-2-isocephem-4-carboxylic acid ##STR170## A.[2-(N-t-Butoxycarbonylaminomethyl)-1-cyclohexen-1-yl]-acetic acid

A solution ofα-[2-(t-butoxycarbonylaminomethyl)-1,4-cyclohexadienyl]-acetic acid(1.33 g., 5 mmoles) in 3% ammonium hydroxide (10 ml.) was hydrogenatedat 40 psi with palladium on charcoal (10%, 0.2 g.). A theoretical amountof hydrogen was taken up in 3 hours. The catalyst was removed and thefiltrate was acidified to pH 2 with dil. HCl and extracted with ethylacetate (2 × 50 ml.). The combined extracts were washed with water (20ml.), dried with Na₂ SO₄ and evaporated under reduced pressure to affordan oil (1.34 g.) which solidified on standing for several days.Recrystallization from n-hexane - ethyl acetate gave 1.2 g. titleproduct as colorless prisms melting at 118°-119° C.

Ir: ν _(max) ^(nujol) 3450, 1730, 1660, 1510 cm.sup.⁻¹.

Nmr: δ _(ppm) ^(CDCl).spsb.3 1.58 (9H, s, t-butyl-H), 1.50 - 1.90 (4H,m, -CH₂ -), 1.90 - 2.20 (4H, m, allylic methylene-H), 3.18 (2H, s, CH₂-CO), 3.78 (2H, d, 6 Hz, CH₂ -N), 5.00 (1H, br-s, NH), 8.98 (1H, br-s,COOH).

Anal. Calc'd. for C₁₄ H₂₃ NO₄ : C, 62.43; H, 8.61; N, 5.20 Found: C,62.12; H, 8.77; N, 5.37.

B. 7β-[α-(2-t-Butoxycarbonylaminomethyl-1-cyclohexenyl)acetamido]-3-(1-methyltetrazol-5-ylthiomethyl)-Δ³-0-2-isocephem-4-carboxylic acid

To a stirred solution of equimolar amounts of[2-(N-t-butoxycarbonylaminomethyl)-1-cyclohexen-1-yl] acetic acid and2,4-dinitrophenol in ethyl acetate is added an equimolar amount ofN,N'-dicyclohexylcarbodiimide. The reaction mixture is stirred for 1hour at room temperature and the precipitated dicyclohexylurea isfiltered off. The filtrate is cooled to 5° C. and poured into a coldsolution of 7β-amino-3-(1-methyltetrazol-5-ylthiomethyl)-Δ³-0-2-isocephem-4-carboxylic acid and excess triethylamine in 50% aqueousTHF. The mixture is stirred overnight at room temperature and washedwith ether. The aqueous layer is acidified with dilute HCl toprecipitate the title product.

C.7β-[α-aminomethyl-1-cyclohexenyl)acetamido]-3-(1-methyltetrazol-5-ylthiomethyl)-Δ³-0-2-isocephem-4-carboxylic acid

A solution of7β-[α-(2-t-butoxycarbonylaminomethyl-1-cyclohexenyl)acetamido]-3-(1-methyltetrazol-5-ylthiomethyl)-Δ³-0-2-isocephem-4-carboxylic acid in trifluoroacetic acid is stirred at0° C. for 1.5 hours. The mixture is diluted with ether to separate thetrifluoroacetate salt which is dissolved in water and neutralized togive the title product.

EXAMPLE 44

When the p-nitrobenzyl 7β-amino-3-(1-methyltetrazol-5-ylthiomethyl)-Δ³-0-2-isocephem-4-carboxylate of Example 15 is replaced by an equimolarweight of p-nitrobenzyl 7β-amino-3-(1,2,3-triazol-5-ylthiomethyl)-Δ³-0-2-isocephem-4-carboxylate, p-nitrobenzyl7β-amino-3-(1-carboxymethyl-1,2,3,4-tetrazol-5-ylthiomethyl)-Δ.sup.3-0-2-isocephem-4-carboxylate and p-nitrobenzyl7β-amino-3-(1-carboxyethyl-1,2,3,4-tetrazol-5-ylthiomethyl)-Δ.sup.3-0-2-isocephem-4-carboxylate, respectively, there are produced aftercatalytic hydrogenation as in the procedure of Example 207β-(2-thienylacetamido)-3-(1,2,3-triazol-5-ylthiomethyl)-Δ.sup.3-0-2-isocephem-4-carboxylic acid,7β-(2-thienylacetamido)-3-(1-carboxymethyl-1,2,3,4-tetrazol-5-ylthiomethyl)-Δ³-0-2-isocephem-4-carboxylic acid and7β-(2-thienylacetamido)-3-(1-carboxyethyl-1,2,3,4-tetrazol-5-ylthiomethyl)-Δ³-0-2-isocephem-4-carboxylic acid, respectively.

EXAMPLE 45

When the 7β-amino-3-(1-methyltetrazol-5-ylthiomethyl)-Δ³-0-2-isocephem-4-carboxylic acid (or ester or salt thereof) in theprocedures of Examples 17, 18, 24 and 25-43 is replaced by an equimolaramount of 7β-amino-3-(1,2,3-triazol-5-ylthiomethyl)-Δ³-0-2-isocephem-4-carboxylic acid,7β-amino-3-(2-methyl-1,3,4-thiadiazol-5-ylthiomethyl)-Δ³-0-2-isocephem-4-carboxylic acid,7β-amino-3-(2-methyl-1,3,4-oxadiazol-5-ylthiomethyl)-Δ³-0-2-isocephem-4-carboxylic acid,7β-amino-3-(1-carboxymethyltetrazol-5-ylthiomethyl)-Δ³-0-2-isocephem-4-carboxylic acid or7β-amino-3-(1-carboxyethyltetrazol-5-ylthiomethyl)-Δ³-0-2-isocephem-4-carboxylic acid (or an ester or salt thereof, anyreactive functional groups other than the 7- amino group being suitablyprotected if necessary), respectively, there are produced (after anynecessary de-blocking of functional protecting groups) the corresponding7β-acylamino carboxylic acids of each of the above-named nuclei.

EXAMPLE 46

When the 7β-amino-3-(1-methyltetrazol-5-ylthiomethyl)-Δ³-0-2-isocephem-4-carboxylic acid (or ester or salt thereof) in theprocedures of Examples 15, 17, 24, and 25-43 is replaced by an equimolaramount of each of the 7β-amino esters of Example 10 (any reactivefunctional groups other than the 7-amino groups being suitably protectedif necessary), there are produced (after any necessary de-blocking offunctional protecting groups of the p-nitrobenzyl ester group) thecorresponding 7β-acylamino carboxylic acids of each of the 7β-aminoesters.

EXAMPLE 47

When the α-amino products of Examples 24 and 37-38, 45 and 46 arereacted with acetone according to the procedure of U.S. Pat. No.3,303,193, there are obtained the corresponding 0-2-isocephemderivatives of the formula ##STR171## where R^(a) is as defined in theabove-mentioned examples and Z represents the appropriate alkyl, aryl,aralkyl or heterocyclic group of the selected nucleus, orpharmaceutically acceptable salts thereof.

EXAMPLE 48

When the α-amino products of Examples 24, 37-38, 45 or 46 are reactedwith dicyanogen or cyanogen bromide or cyanogen chloride according tothe procedure disclosed in U.S. Pat. No. 3,796,709, the correspondingα-cyanamino products are obtained.

EXAMPLE 49

When the α-amino products of Examples 24, 37-38, 45 or 46 are reactedwith a triethylamine --SO₃ complex according to the procedure of U.S.Pat. No. 3,381,001, the corresponding α-sulfoamino products areobtained.

EXAMPLE 50

When the α-amino products of Examples 24, 37-38, 45 or 46 are reactedwith 1-methyl-1-nitrosobiuret or a1-methyl-5-(lower)alkyl-1-nitrosobiuret according to the procedure ofU.S. Pat. No. 3,483,188, there are produced the compounds of the generalformula ##STR172## where R² is hydrogen or (lower)alkyl, Z representsthe appropriate alkyl, aryl, aralkyl or heterocyclic group of theselected nucleus and R^(a) is as defined in the above-mentionedexamples.

EXAMPLE 51

When the 7-amino intermediates in the acylation procedures of Examples15, 16, 17, 19, 24 and 25-46 are replaced by the correspondingpivaloyloxymethyl, acetoxymethyl, methoxymethyl, acetonyl and phenacylesters, respectively, and the ester group of the 7-acylamido product isnot removed, there are obtained the corresponding pivaloyloxymethyl,acetoxymethyl, methoxymethyl, acetonyl and phenacyl esters,respectively, of the 7-acylamido end-products.

EXAMPLE 52

7β-phenoxyacetamido-3-(1-methyltetrazol-5-ylthiomethyl)-Δ³-0-2-isocephem-4-carboxylic acid (alternate process)

A mixture of benzyl 7β-phenoxyacetamido-3-methylsulfonyloxymethyl-Δ³-0-2-isocephem-4-carboxylate (0.22 g., 0.43 mmole; prepared according topreparation 4 above), triethylamine (0.07 ml., 0.5 mmole) and1-methyltetrazole thiol (0.05 g., 0.5 mmole) in methylene chloride (50ml.) was stirred at room temperature for 16 hours, washed with 10% HCl(20 ml.) and brine (2 × 50 ml.), dried and concentrated to give asemi-solid (0.24 g.) which was purified by column chromatography toafford 0.15 g. of pure benzyl7β-phenoxyacetamido-3-(1-methyltetrazol-5-ylthiomethyl)-Δ³-0-2-isocephem-4-carboxylate as a white semi-solid. The product wasshown by IR and NMR to be identical with the product of Example 18.

Anal. Calc'd. for C₂₅ H₂₄ N₆ O₆ S: C, 55.97; H, 4.50; N, 15.66. Found:C, 55.89; H, 4.55; N, 15.34.

The benzyl ester is subjected to catalytic hydrogenation following theprocedure of Example 23 to give the title product.

We claim:
 1. The compound of the formula ##STR173## wherein R" ishydrogen or an easily cleavable ester carboxyl-protecting group and Zrepresents a 5- or 6-membered heterocyclic ring containing N, O or S,said heterocyclic ring being optionally substituted by one or twosubstituents selected from the group consisting of halo, C₁ -C₄ alkyl,C₁ -C₄ alkoxy, cyano, carboxyl, amino, nitro, C₃ -C₄ cycloalkyl, C₂ -C₄alkenyl, trifluoromethyl, hydroxy, hydroxymethyl, C₁ -C₄ alkylthio, C₁-C₄ alkylamino, di(C₁ -C₄ alkyl)amino, mercapto, phenyl, benzyl,alkoxyalkyl of up to 4 carbons and -(CH₂)_(n) COOH in which n is aninteger of 1 to 4, or carboxylic acid or acid addition salts thereof. 2.An acid, ester or salt of claim 1 wherein Z represents a heterocyclicradical selected from the group consisting of thienyl, furyl, pyrazolyl,imidazolyl, isoimidazolyl, triazolyl, tetrazolyl, thiazolyl,thiadiazolyl, oxazolyl, oxadiazolyl, isothiazolyl, isoxazolyl, pyridyl,pryidazinyl, pyrazinyl, pyrimidinyl and triazinyl, said heterocyclicradical being optionally substituted by one or two substituents selectedfrom the group consisting of halo, C₁ -C₄ alkyl, C₁ -C₄ alkoxy, cyano,carboxyl, amino, nitro, C₃ -C₄ cycloalkyl, C₂ -C₄ alkenyl,trifluoromethyl, hydroxy, hydroxymethyl, C₁ -C₄ alkylthio, C₁ -C₄alkylamino, di(C₁ -C₄ alkyl)amino, mercapto, phenyl, benzyl, alkoxyalkylof up to 4 carbons and --(CH₂)_(n) COOH in which n is an integer of 1 to4.
 3. An acid, ester or salt of claim 2 wherein Z represents a triazole,thiadiazole, oxadiazole or tetrazole radical, said radical beingoptionally substituted by one or two substituents selected from thegroup consisting of halo, C₁ -C₄ alkyl, C₁ -C₄ alkoxy, cyano, carboxyl,amino, nitro, C₃ -C₄ cycloalkyl, C₂ -C₄ alkenyl, trifluoromethyl,hydroxy, hydroxymethyl, C₁ -C₄ alkylthio, C₁ -C₄ alkylamino, di(C₁ -C₄alkyl)amino, phenyl, benzyl, mercapto, alkoxyalkyl of up to 4 carbonsand --(CH₂)_(n) COOH in which n is an integer of 1 to
 4. 4. A compoundof claim 1 wherein R" is hydrogen, or a carboxylic acid or acid additionsalt thereof.
 5. A compound of claim 1 wherein R" is hydrogen.
 6. Acompound of claim 2 wherein R" is hydrogen, or a carboxylic acid or acidaddition salt thereof.
 7. A compound of claim 2 wherein R" is hydrogen.8. A compound of claim 3 wherein R" is hydrogen, or a carboxylic acid oracid addition salt thereof.
 9. A compound of claim 3 wherein R" ishydrogen.
 10. The compound of the formula ##STR174## wherein R" ishydrogen or an easily cleavable ester carboxyl-protecting group and Zrepresents 1,2,3-triazol-5-yl, 2-methyl-1,3,4-thiadiazol-5-yl,2-methyl-1,3,4-oxadiazol-5-yl, 1-N-methyltetrazol-5-yl,1-carboxymethyltetrazol-5-yl or 1-carboxyethyltetrazol-5-yl orcarboxylic acid or acid addition salts thereof.
 11. An acid of claim 10wherein R" is hydrogen, or a carboxylic acid or acid addition saltthereof.
 12. An ester of claim 10 wherein R" is benzyhydryl, benzyl,p-nitrobenzyl, p-methoxybenzyl, trichloroethyl, trimethylsilyl,phenacyl, acetonyl, (lower)alkyl, triphenylmethyl, methoxymethyl,indanyl, phthalidyl, pivaloxymethyl or acetoxymethyl, or an acidaddition salt thereof.
 13. The acid of claim 10 wherein Z is1,2,3-triazol-5-yl.
 14. The acid of claim 10 wherein Z is2-methyl-1,3,4-thiadiazol-5-yl.
 15. The acid of claim 10 wherein Z is2-methyl-1,3,4-oxadiazol-5-yl.
 16. The acid of claim 10 wherein Z is1-N-methyltetrazol-5-yl.
 17. The acid of claim 10 wherein Z is1-carboxymethyltetrazol-5-yl.
 18. The acid of claim 10 wherein Z is1-carboxyethyltetrazol-5-yl.
 19. The ester of claim 10 namedp-nitrobenzyl 7β-amino-3-(1-methyltetrazol-5-ylthiomethyl)-Δ³-0-2-isocephem-4-carboxylate.
 20. The ester of claim 10 named benzyl7β-amino-3-(1-methyltetrazol-5-ylthiomethyl)-Δ³-0-2-isocephem-4-carboxylate.
 21. The ester of claim 10 namedp-nitrobenzyl 7β-amino-3-(2-methyl-1,3,4-thiadiazol-5-ylthiomethyl)-Δ³-0-2-isocephem-4-carboxylate.
 22. The ester of claim 10 namedp-nitrobenzyl 7β-amino-3-(2-methyl-1,3,4-oxadiazol-5-ylthiomethyl)-Δ³-0-2-isocephem-4-carboxylate.
 23. An acid of claim 10 wherein R" ishydrogen.